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Insulin crystals.

Biotechnology is a field of biology that involves the use of living things in engineering, technology, medicine, etc.. Modern use of the term refers to genetic engineering as well as cell- and tissue culture technologies. However, the concept encompasses a wider range and history of procedures for modifying living organisms according to human purposes, going back to domestication of animals, cultivation of plants and "improvements" to these through breeding programs that employ artificial selection and hybridization. By comparison to biotechnology, bioengineering is generally thought of as a related field with its emphasis more on mechanical and higher systems approaches to interfacing with and exploiting living things. United Nations Convention on Biological Diversity defines biotechnology as:[1]

"Any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use."

Biotechnology draws on the pure biological sciences (genetics, microbiology, animal cell culture, molecular biology, biochemistry, embryology, cell biology) and in many instances is also dependent on knowledge and methods from outside the sphere of biology (chemical engineering, bioprocess engineering, information technology, biorobotics). Conversely, modern biological sciences (including even concepts such as molecular ecology) are intimately entwined and dependent on the methods developed through biotechnology and what is commonly thought of as the life sciences industry.

Contents

History

Brewing was an early application of biotechnology

Although not normally thought of as biotechnology, agriculture clearly fits the broad definition of "using a biotechnological system to make products" such that the cultivation of plants may be viewed as the earliest biotechnological enterprise. Agriculture has been theorized to have become the dominant way of producing food since the Neolithic Revolution. The processes and methods of agriculture have been refined by other mechanical and biological sciences since its inception. Through early biotechnology, farmers were able to select the best suited and highest-yield crops to produce enough food to support a growing population. Other uses of biotechnology were required as crops and fields became increasingly large and difficult to maintain. Specific organisms and organism by-products were used to fertilize, restore nitrogen, and control pests. Throughout the use of agriculture, farmers have inadvertently altered the genetics of their crops through introducing them to new environments and breeding them with other plants—one of the first forms of biotechnology. Cultures such as those in Mesopotamia, Egypt, and India developed the process of brewing beer. It is still done by the same basic method of using malted grains (containing enzymes) to convert starch from grains into sugar and then adding specific yeasts to produce beer. In this process the carbohydrates in the grains were broken down into alcohols such as ethanol. Ancient Indians also used the juices of the plant Ephedra vulgaris and used to call it Soma.[citation needed] Later other cultures produced the process of Lactic acid fermentation which allowed the fermentation and preservation of other forms of food. Fermentation was also used in this time period to produce leavened bread. Although the process of fermentation was not fully understood until Pasteur’s work in 1857, it is still the first use of biotechnology to convert a food source into another form.

Combinations of plants and other organisms were used as medications in many early civilizations. Since as early as 200 BC, people began to use disabled or minute amounts of infectious agents to immunize themselves against infections. These and similar processes have been refined in modern medicine and have led to many developments such as antibiotics, vaccines, and other methods of fighting sickness.[citation needed]

In the early twentieth century scientists gained a greater understanding of microbiology and explored ways of manufacturing specific products. In 1917, Chaim Weizmann first used a pure microbiological culture in an industrial process, that of manufacturing corn starch using Clostridium acetobutylicum, to produce acetone, which the United Kingdom desperately needed to manufacture explosives during World War I.[2]

The field of modern biotechnology is thought to have largely begun on June 16, 1980, when the United States Supreme Court ruled that a genetically-modified microorganism could be patented in the case of Diamond v. Chakrabarty.[3] Indian-born Ananda Chakrabarty, working for General Electric, had developed a bacterium (derived from the Pseudomonas genus) capable of breaking down crude oil, which he proposed to use in treating oil spills.

Revenue in the industry is expected to grow by 12.9% in 2008. Another factor influencing the biotechnology sector's success is improved intellectual property rights legislation—and enforcement—worldwide, as well as strengthened demand for medical and pharmaceutical products to cope with an ageing, and ailing, U.S. population.[4]

Rising demand for biofuels is expected to be good news for the biotechnology sector, with the Department of Energy estimating ethanol usage could reduce U.S. petroleum-derived fuel consumption by up to 30% by 2030. The biotechnology sector has allowed the U.S. farming industry to rapidly increase its supply of corn and soybeans—the main inputs into biofuels—by developing genetically-modified seeds which are resistant to pests and drought. By boosting farm productivity, biotechnology plays a crucial role in ensuring that biofuel production targets are met.[5]

Applications

A rose plant that began as cells grown in a tissue culture

Biotechnology has applications in four major industrial areas, including health care (medical), crop production and agriculture, non food (industrial) uses of crops and other products (e.g. biodegradable plastics, vegetable oil, biofuels), and environmental uses.

For example, one application of biotechnology is the directed use of organisms for the manufacture of organic products (examples include beer and milk products). Another example is using naturally present bacteria by the mining industry in bioleaching. Biotechnology is also used to recycle, treat waste, clean up sites contaminated by industrial activities (bioremediation), and also to produce biological weapons.

A series of derived terms have been coined to identify several branches of biotechnology, for example:

  • Bioinformatics is an interdisciplinary field which addresses biological problems using computational techniques, and makes the rapid organization and analysis of biological data possible. The field may also be referred to as computational biology, and can be defined as, "conceptualizing biology in terms of molecules and then applying informatics techniques to understand and organize the information associated with these molecules, on a large scale."[6] Bioinformatics plays a key role in various areas, such as functional genomics, structural genomics, and proteomics, and forms a key component in the biotechnology and pharmaceutical sector.
  • Blue biotechnology is a term that has been used to describe the marine and aquatic applications of biotechnology, but its use is relatively rare.
  • Green biotechnology is biotechnology applied to agricultural processes. An example would be the selection and domestication of plants via micropropagation. Another example is the designing of transgenic plants to grow under specific environments in the presence (or absence) of chemicals. One hope is that green biotechnology might produce more environmentally friendly solutions than traditional industrial agriculture. An example of this is the engineering of a plant to express a pesticide, thereby ending the need of external application of pesticides. An example of this would be Bt corn. Whether or not green biotechnology products such as this are ultimately more environmentally friendly is a topic of considerable debate.
  • Red biotechnology is applied to medical processes. Some examples are the designing of organisms to produce antibiotics, and the engineering of genetic cures through genetic manipulation.
  • White biotechnology, also known as industrial biotechnology, is biotechnology applied to industrial processes. An example is the designing of an organism to produce a useful chemical. Another example is the using of enzymes as industrial catalysts to either produce valuable chemicals or destroy hazardous/polluting chemicals. White biotechnology tends to consume less in resources than traditional processes used to produce industrial goods.[citation needed] The investment and economic output of all of these types of applied biotechnologies is termed as bioeconomy.

Medicine

In medicine, modern biotechnology finds promising applications in such areas as

Pharmacogenomics

DNA Microarray chip – Some can do as many as a million blood tests at once

Pharmacogenomics is the study of how the genetic inheritance of an individual affects his/her body’s response to drugs. It is a coined word derived from the words “pharmacology” and “genomics”. It is hence the study of the relationship between pharmaceuticals and genetics. The vision of pharmacogenomics is to be able to design and produce drugs that are adapted to each person’s genetic makeup.[7]

Pharmacogenomics results in the following benefits:[7]

  1. Development of tailor-made medicines. Using pharmacogenomics, pharmaceutical companies can create drugs based on the proteins, enzymes and RNA molecules that are associated with specific genes and diseases. These tailor-made drugs promise not only to maximize therapeutic effects but also to decrease damage to nearby healthy cells.
  2. More accurate methods of determining appropriate drug dosages. Knowing a patient’s genetics will enable doctors to determine how well his/ her body can process and metabolize a medicine. This will maximize the value of the medicine and decrease the likelihood of overdose.
  3. Improvements in the drug discovery and approval process. The discovery of potential therapies will be made easier using genome targets. Genes have been associated with numerous diseases and disorders. With modern biotechnology, these genes can be used as targets for the development of effective new therapies, which could significantly shorten the drug discovery process.
  4. Better vaccines. Safer vaccines can be designed and produced by organisms transformed by means of genetic engineering. These vaccines will elicit the immune response without the attendant risks of infection. They will be inexpensive, stable, easy to store, and capable of being engineered to carry several strains of pathogen at once.

Pharmaceutical products

Computer-generated image of insulin hexamers highlighting the threefold symmetry, the zinc ions holding it together, and the histidine residues involved in zinc binding.

Most traditional pharmaceutical drugs are relatively simple molecules that have been found primarily through trial and error to treat the symptoms of a disease or illness.[citation needed] Biopharmaceuticals are large biological molecules known as proteins and these usually target the underlying mechanisms and pathways of a malady (but not always, as is the case with using insulin to treat type 1 diabetes mellitus, as that treatment merely addresses the symptoms of the disease, not the underlying cause which is autoimmunity); it is a relatively young industry. They can deal with targets in humans that may not be accessible with traditional medicines. A patient typically is dosed with a small molecule via a tablet while a large molecule is typically injected.

Small molecules are manufactured by chemistry but larger molecules are created by living cells such as those found in the human body: for example, bacteria cells, yeast cells, animal or plant cells.

Modern biotechnology is often associated with the use of genetically altered microorganisms such as E. coli or yeast for the production of substances like synthetic insulin or antibiotics. It can also refer to transgenic animals or transgenic plants, such as Bt corn. Genetically altered mammalian cells, such as Chinese Hamster Ovary (CHO) cells, are also used to manufacture certain pharmaceuticals. Another promising new biotechnology application is the development of plant-made pharmaceuticals.

Biotechnology is also commonly associated with landmark breakthroughs in new medical therapies to treat hepatitis B, hepatitis C, cancers, arthritis, haemophilia, bone fractures, multiple sclerosis, and cardiovascular disorders. The biotechnology industry has also been instrumental in developing molecular diagnostic devices that can be used to define the target patient population for a given biopharmaceutical. Herceptin, for example, was the first drug approved for use with a matching diagnostic test and is used to treat breast cancer in women whose cancer cells express the protein HER2.

Modern biotechnology can be used to manufacture existing medicines relatively easily and cheaply. The first genetically engineered products were medicines designed to treat human diseases. To cite one example, in 1978 Genentech developed synthetic humanized insulin by joining its gene with a plasmid vector inserted into the bacterium Escherichia coli. Insulin, widely used for the treatment of diabetes, was previously extracted from the pancreas of abattoir animals (cattle and/or pigs). The resulting genetically engineered bacterium enabled the production of vast quantities of synthetic human insulin at relatively low cost.[8] According to a 2003 study undertaken by the International Diabetes Federation (IDF) on the access to and availability of insulin in its member countries, synthetic 'human' insulin is considerably more expensive in most countries where both synthetic 'human' and animal insulin are commercially available: e.g. within European countries the average price of synthetic 'human' insulin was twice as high as the price of pork insulin.[9] Yet in its position statement, the IDF writes that "there is no overwhelming evidence to prefer one species of insulin over another" and "[modern, highly-purified] animal insulins remain a perfectly acceptable alternative.[10]

Modern biotechnology has evolved, making it possible to produce more easily and relatively cheaply human growth hormone, clotting factors for hemophiliacs, fertility drugs, erythropoietin and other drugs.[11] Most drugs today are based on about 500 molecular targets. Genomic knowledge of the genes involved in diseases, disease pathways, and drug-response sites are expected to lead to the discovery of thousands more new targets.[11]

Genetic testing

Genetic testing involves the direct examination of the DNA molecule itself. A scientist scans a patient’s DNA sample for mutated sequences.

There are two major types of gene tests. In the first type, a researcher may design short pieces of DNA (“probes”) whose sequences are complementary to the mutated sequences. These probes will seek their complement among the base pairs of an individual’s genome. If the mutated sequence is present in the patient’s genome, the probe will bind to it and flag the mutation. In the second type, a researcher may conduct the gene test by comparing the sequence of DNA bases in a patient’s gene to disease in healthy individuals or their progeny.

Genetic testing is now used for:

  • Carrier screening, or the identification of unaffected individuals who carry one copy of a gene for a disease that requires two copies for the disease to manifest;
  • Confirmational diagnosis of symptomatic individuals;
  • Determining sex;
  • Forensic/identity testing;
  • Newborn screening;
  • Prenatal diagnostic screening;
  • Presymptomatic testing for estimating the risk of developing adult-onset cancers;
  • Presymptomatic testing for predicting adult-onset disorders.

Some genetic tests are already available, although most of them are used in developed countries. The tests currently available can detect mutations associated with rare genetic disorders like cystic fibrosis, sickle cell anemia, and Huntington’s disease. Recently, tests have been developed to detect mutation for a handful of more complex conditions such as breast, ovarian, and colon cancers. However, gene tests may not detect every mutation associated with a particular condition because many are as yet undiscovered, and the ones they do detect may present different risks to different people and populations.[11]

Controversial questions
The bacterium Escherichia coli is routinely genetically engineered.

The absence of privacy and anti-discrimination legal protections in most countries can lead to discrimination in employment or insurance or other misuse of personal genetic information. This raises questions such as whether genetic privacy is different from medical privacy.[12]

  1. Reproductive issues. These include the use of genetic information in reproductive decision-making and the possibility of genetically altering reproductive cells that may be passed on to future generations. For example, germline therapy forever changes the genetic make-up of an individual’s descendants. Thus, any error in technology or judgment may have far-reaching consequences. Ethical issues like designer babies and human cloning have also given rise to controversies between and among scientists and bioethicists, especially in the light of past abuses with eugenics.
  2. Clinical issues. These center on the capabilities and limitations of doctors and other health-service providers, people identified with genetic conditions, and the general public in dealing with genetic information.
  3. Effects on social institutions. Genetic tests reveal information about individuals and their families. Thus, test results can affect the dynamics within social institutions, particularly the family.
  4. Conceptual and philosophical implications regarding human responsibility, free will vis-à-vis genetic determinism, and the concepts of health and disease.

Gene therapy

Gene therapy using an Adenovirus vector. A new gene is inserted into an adenovirus vector, which is used to introduce the modified DNA into a human cell. If the treatment is successful, the new gene will make a functional protein.

Gene therapy may be used for treating, or even curing, genetic and acquired diseases like cancer and AIDS by using normal genes to supplement or replace defective genes or to bolster a normal function such as immunity. It can be used to target somatic (i.e., body) or gametes (i.e., egg and sperm) cells. In somatic gene therapy, the genome of the recipient is changed, but this change is not passed along to the next generation. In contrast, in germline gene therapy, the egg and sperm cells of the parents are changed for the purpose of passing on the changes to their offspring.

There are basically two ways of implementing a gene therapy treatment:

  1. Ex vivo, which means “outside the body” – Cells from the patient’s blood or bone marrow are removed and grown in the laboratory. They are then exposed to a virus carrying the desired gene. The virus enters the cells, and the desired gene becomes part of the DNA of the cells. The cells are allowed to grow in the laboratory before being returned to the patient by injection into a vein.
  2. In vivo, which means “inside the body” – No cells are removed from the patient’s body. Instead, vectors are used to deliver the desired gene to cells in the patient’s body.

As of June 2001, more than 500 clinical gene-therapy trials involving about 3,500 patients have been identified worldwide. Around 78% of these are in the United States, with Europe having 18%. These trials focus on various types of cancer, although other multigenic diseases are being studied as well. Recently, two children born with severe combined immunodeficiency disorder (“SCID”) were reported to have been cured after being given genetically engineered cells.

Gene therapy faces many obstacles before it can become a practical approach for treating disease.[13] At least four of these obstacles are as follows:

  1. Gene delivery tools. Genes are inserted into the body using gene carriers called vectors. The most common vectors now are viruses, which have evolved a way of encapsulating and delivering their genes to human cells in a pathogenic manner. Scientists manipulate the genome of the virus by removing the disease-causing genes and inserting the therapeutic genes. However, while viruses are effective, they can introduce problems like toxicity, immune and inflammatory responses, and gene control and targeting issues. In addition, in order for gene therapy to provide permanent therapeutic effects, the introduced gene needs to be integrated within the host cell's genome. Some viral vectors effect this in a random fashion, which can introduce other problems such as disruption of an endogenous host gene.
  2. High costs. Since gene therapy is relatively new and at an experimental stage, it is an expensive treatment to undertake. This explains why current studies are focused on illnesses commonly found in developed countries, where more people can afford to pay for treatment. It may take decades before developing countries can take advantage of this technology.
  3. Limited knowledge of the functions of genes. Scientists currently know the functions of only a few genes. Hence, gene therapy can address only some genes that cause a particular disease. Worse, it is not known exactly whether genes have more than one function, which creates uncertainty as to whether replacing such genes is indeed desirable.
  4. Multigene disorders and effect of environment. Most genetic disorders involve more than one gene. Moreover, most diseases involve the interaction of several genes and the environment. For example, many people with cancer not only inherit the disease gene for the disorder, but may have also failed to inherit specific tumor suppressor genes. Diet, exercise, smoking and other environmental factors may have also contributed to their disease.

Human Genome Project

DNA Replication image from the Human Genome Project (HGP)

The Human Genome Project is an initiative of the U.S. Department of Energy (“DOE”) that aims to generate a high-quality reference sequence for the entire human genome and identify all the human genes.

The DOE and its predecessor agencies were assigned by the U.S. Congress to develop new energy resources and technologies and to pursue a deeper understanding of potential health and environmental risks posed by their production and use. In 1986, the DOE announced its Human Genome Initiative. Shortly thereafter, the DOE and National Institutes of Health developed a plan for a joint Human Genome Project (“HGP”), which officially began in 1990.

The HGP was originally planned to last 15 years. However, rapid technological advances and worldwide participation accelerated the completion date to 2003 (making it a 13 year project). Already it has enabled gene hunters to pinpoint genes associated with more than 30 disorders.[14]

Cloning

Cloning involves the removal of the nucleus from one cell and its placement in an unfertilized egg cell whose nucleus has either been deactivated or removed.

There are two types of cloning:

  1. Reproductive cloning. After a few divisions, the egg cell is placed into a uterus where it is allowed to develop into a fetus that is genetically identical to the donor of the original nucleus.
  2. Therapeutic cloning.[15] The egg is placed into a Petri dish where it develops into embryonic stem cells, which have shown potentials for treating several ailments.[16]

In February 1997, cloning became the focus of media attention when Ian Wilmut and his colleagues at the Roslin Institute announced the successful cloning of a sheep, named Dolly, from the mammary glands of an adult female. The cloning of Dolly made it apparent to many that the techniques used to produce her could someday be used to clone human beings.[17] This stirred a lot of controversy because of its ethical implications.

Agriculture

Responsible biotechnology is not the enemy; starvation is. Without adequate food supplies at affordable prices, we cannot expect world health or peace.
Jimmy Carter, Former President of the United States, 11 Jul 1997.[18]

Crop yield

Using the techniques of modern biotechnology, one or two genes(Smartstax from Monsanto in collaboration with Dow AgroSciences will use 8, starting in 2010) may be transferred to a highly developed crop variety to impart a new character that would increase its yield.[19] However, while increases in crop yield are the most obvious applications of modern biotechnology in agriculture, it is also the most difficult one. Current genetic engineering techniques work best for effects that are controlled by a single gene. Many of the genetic characteristics associated with yield (e.g., enhanced growth) are controlled by a large number of genes, each of which has a minimal effect on the overall yield.[20] There is, therefore, much scientific work to be done in this area.

Reduced vulnerability of crops to environmental stresses

Crops containing genes that will enable them to withstand biotic and abiotic stresses may be developed. For example, drought and excessively salty soil are two important limiting factors in crop productivity. Biotechnologists are studying plants that can cope with these extreme conditions in the hope of finding the genes that enable them to do so and eventually transferring these genes to the more desirable crops. One of the latest developments is the identification of a plant gene, At-DBF2, from Arabidopsis thaliana, a tiny weed that is often used for plant research because it is very easy to grow and its genetic code is well mapped out. When this gene was inserted into tomato and tobacco cells (see RNA interference), the cells were able to withstand environmental stresses like salt, drought, cold and heat, far more than ordinary cells. If these preliminary results prove successful in larger trials, then At-DBF2 genes can help in engineering crops that can better withstand harsh environments.[21] Researchers have also created transgenic rice plants that are resistant to rice yellow mottle virus (RYMV). In Africa, this virus destroys majority of the rice crops and makes the surviving plants more susceptible to fungal infections.[22]

Increased nutritional qualities

Proteins in foods may be modified to increase their nutritional qualities. Proteins in legumes and cereals may be transformed to provide the amino acids needed by human beings for a balanced diet.[20] A good example is the work of Professors Ingo Potrykus and Peter Beyer on the so-called Golden rice (discussed below).

Improved taste, texture or appearance of food

Modern biotechnology can be used to slow down the process of spoilage so that fruit can ripen longer on the plant and then be transported to the consumer with a still reasonable shelf life. This alters the taste, texture and appearance of the fruit. More importantly, it could expand the market for farmers in developing countries due to the reduction in spoilage. However, there is sometimes a lack of understanding by researchers in developed countries about the actual needs of prospective beneficiaries in developing countries. For example, engineering soybeans to resist spoilage makes them less suitable for producing tempeh which is a significant source of protein that depends on fermentation. The use of modified soybeans results in a lumpy texture that is less palatable and less convenient when cooking.

The first genetically modified food product was a tomato which was transformed to delay its ripening.[23] Researchers in Indonesia, Malaysia, Thailand, Philippines and Vietnam are currently working on delayed-ripening papaya in collaboration with the University of Nottingham and Zeneca.[24]

Biotechnology in cheese production:[25] enzymes produced by micro-organisms provide an alternative to animal rennet – a cheese coagulant – and an alternative supply for cheese makers. This also eliminates possible public concerns with animal-derived material, although there are currently no plans to develop synthetic milk, thus making this argument less compelling. Enzymes offer an animal-friendly alternative to animal rennet. While providing comparable quality, they are theoretically also less expensive.

About 85 million tons of wheat flour is used every year to bake bread.[26] By adding an enzyme called maltogenic amylase to the flour, bread stays fresher longer. Assuming that 10–15% of bread is thrown away as stale, if it could be made to stay fresh another 5–7 days then perhaps 2 million tons of flour per year would be saved. Other enzymes can cause bread to expand to make a lighter loaf, or alter the loaf in a range of ways.

Reduced dependence on fertilizers, pesticides and other agrochemicals

Most of the current commercial applications of modern biotechnology in agriculture are on reducing the dependence of farmers on agrochemicals. For example, Bacillus thuringiensis (Bt) is a soil bacterium that produces a protein with insecticidal qualities. Traditionally, a fermentation process has been used to produce an insecticidal spray from these bacteria. In this form, the Bt toxin occurs as an inactive protoxin, which requires digestion by an insect to be effective. There are several Bt toxins and each one is specific to certain target insects. Crop plants have now been engineered to contain and express the genes for Bt toxin, which they produce in its active form. When a susceptible insect ingests the transgenic crop cultivar expressing the Bt protein, it stops feeding and soon thereafter dies as a result of the Bt toxin binding to its gut wall. Bt corn is now commercially available in a number of countries to control corn borer (a lepidopteran insect), which is otherwise controlled by spraying (a more difficult process).

Crops have also been genetically engineered to acquire tolerance to broad-spectrum herbicide. The lack of herbicides with broad-spectrum activity and no crop injury was a consistent limitation in crop weed management. Multiple applications of numerous herbicides were routinely used to control a wide range of weed species detrimental to agronomic crops. Weed management tended to rely on preemergence—that is, herbicide applications were sprayed in response to expected weed infestations rather than in response to actual weeds present. Mechanical cultivation and hand weeding were often necessary to control weeds not controlled by herbicide applications. The introduction of herbicide-tolerant crops has the potential of reducing the number of herbicide active ingredients used for weed management, reducing the number of herbicide applications made during a season, and increasing yield due to improved weed management and less crop injury. Transgenic crops that express tolerance to glyphosate, glufosinate and bromoxynil have been developed. These herbicides can now be sprayed on transgenic crops without inflicting damage on the crops while killing nearby weeds.[27]

From 1996 to 2001, herbicide tolerance was the most dominant trait introduced to commercially available transgenic crops, followed by insect resistance. In 2001, herbicide tolerance deployed in soybean, corn and cotton accounted for 77% of the 626,000 square kilometres planted to transgenic crops; Bt crops accounted for 15%; and "stacked genes" for herbicide tolerance and insect resistance used in both cotton and corn accounted for 8%.[28]

Production of novel substances in crop plants

Biotechnology is being applied for novel uses other than food. For example, oilseed can be modified to produce fatty acids for detergents, substitute fuels and petrochemicals. Potatoes, tomatoes, rice tobacco, lettuce, safflowers, and other plants have been genetically-engineered to produce insulin and certain vaccines. If future clinical trials prove successful, the advantages of edible vaccines would be enormous, especially for developing countries. The transgenic plants may be grown locally and cheaply. Homegrown vaccines would also avoid logistical and economic problems posed by having to transport traditional preparations over long distances and keeping them cold while in transit. And since they are edible, they will not need syringes, which are not only an additional expense in the traditional vaccine preparations but also a source of infections if contaminated.[29] In the case of insulin grown in transgenic plants, it is well-established that the gastrointestinal system breaks the protein down therefore this could not currently be administered as an edible protein. However, it might be produced at significantly lower cost than insulin produced in costly, bioreactors. For example, Calgary, Canada-based SemBioSys Genetics, Inc.[30] reports that its safflower-produced insulin will reduce unit costs by over 25% or more and approximates a reduction in the capital costs associated with building a commercial-scale insulin manufacturing facility of over $100 million, compared to traditional biomanufacturing facilities.[31]

Criticism

There is another side to the agricultural biotechnology issue. It includes increased herbicide usage and resultant herbicide resistance, "super weeds," residues on and in food crops, genetic contamination of non-GM crops which hurt organic and conventional farmers, damage for all wildlife from glyphosate, etc.[32][33]

Biological engineering

Biotechnological engineering or biological engineering is a branch of engineering that focuses on biotechnologies and biological science. It includes different disciplines such as biochemical engineering, biomedical engineering, bio-process engineering, biosystem engineering and so on. Because of the novelty of the field, the definition of a bioengineer is still undefined. However, in general it is an integrated approach of fundamental biological sciences and traditional engineering principles.

Biotechnologist are often employed to scale up bio processes from the laboratory scale to the manufacturing scale. Moreover, as with most engineers, they often deal with management, economic and legal issues. Since patents and regulation (e.g., U.S. Food and Drug Administration regulation in the U.S.) are very important issues for biotech enterprises, bioengineers are often required to have knowledge related to these issues.

The increasing number of biotech enterprises is likely to create a need for bioengineers in the years to come. Many universities throughout the world are now providing programs in bioengineering and biotechnology (as independent programs or specialty programs within more established engineering fields).

Bioremediation and Biodegradation

Biotechnology is being used to engineer and adapt organisms especially microorganisms in an effort to find sustainable ways to clean up contaminated environments. The elimination of a wide range of pollutants and wastes from the environment is an absolute requirement to promote a sustainable development of our society with low environmental impact. Biological processes play a major role in the removal of contaminants and biotechnology is taking advantage of the astonishing catabolic versatility of microorganisms to degrade/convert such compounds. New methodological breakthroughs in sequencing, genomics, proteomics, bioinformatics and imaging are producing vast amounts of information. In the field of Environmental Microbiology, genome-based global studies open a new era providing unprecedented in silico views of metabolic and regulatory networks, as well as clues to the evolution of degradation pathways and to the molecular adaptation strategies to changing environmental conditions. Functional genomic and metagenomic approaches are increasing our understanding of the relative importance of different pathways and regulatory networks to carbon flux in particular environments and for particular compounds and they will certainly accelerate the development of bioremediation technologies and biotransformation processes.[34]

Marine environments are especially vulnerable since oil spills of coastal regions and the open sea are poorly containable and mitigation is difficult. In addition to pollution through human activities, millions of tons of petroleum enter the marine environment every year from natural seepages. Despite its toxicity, a considerable fraction of petroleum oil entering marine systems is eliminated by the hydrocarbon-degrading activities of microbial communities, in particular by a remarkable recently discovered group of specialists, the so-called hydrocarbonoclastic bacteria (HCCB).[35]

Education

In 1988, after prompting from the United States Congress, the National Institute of General Medical Sciences (National Institutes of Health) instituted a funding mechanism for biotechnology training. Universities nationwide compete for these funds to establish Biotechnology Training Programs (BTPs). Each successful application is generally funded for five years then must be competitively renewed. Graduate students in turn compete for acceptance into a BTP; if accepted then stipend, tuition and health insurance support is provided for two or three years during the course of their PhD thesis work. Nineteen institutions offer NIGMS supported BTPs.[36] Biotechnology training is also offered at the undergraduate level and in community colleges.

Notable researchers and individuals

See also

References

  1. ^ "The Convention on Biological Diversity (Article 2. Use of Terms)." United Nations. 1992. Retrieved on February 6, 2008.
  2. ^ Springham, D.; Springham, G.; Moses, V.; Cape, R.E. "Biotechnology: The Science and the Business." Published 1999, Taylor & Francis. p. 1. ISBN 9057024071
  3. ^ "Diamond v. Chakrabarty, 447 U.S. 303 (1980). No. 79-139." United States Supreme Court. June 16, 1980. Retrieved on May 4, 2007.
  4. ^ IBISWorld
  5. ^ The Recession List - Top 10 Industries to Fly and Fl... (ith anincreasing share accounted for by ...), bio-medicine.org
  6. ^ Gerstein, M. "Bioinformatics Introduction." Yale University. Retrieved on May 8, 2007.
  7. ^ a b U.S. Department of Energy Human Genome Program, supra note 6.
  8. ^ W. Bains, Genetic Engineering For Almost Everybody: What Does It Do? What Will It Do? (London: Penguin Books, 1987), 99.
  9. ^ IDF 2003; "Diabetes Atlas,: 2nd ed."; International Diabetes Federation, Brussels, eatlas.idf.org
  10. ^ IDF March 2005; "Position Statement." International Diabetes Federation, Brussels. idf.org
  11. ^ a b c U.S. Department of State International Information Programs, “Frequently Asked Questions About Biotechnology”, USIS Online; available from USinfo.state.gov, accessed 13 September 2007. Cf. C. Feldbaum, “Some History Should Be Repeated”, 295 Science, 8 February 2002, 975.
  12. ^ The National Action Plan on Breast Cancer and U.S. National Institutes of Health-Department of Energy Working Group on the Ethical, Legal and Social Implications (ELSI) have issued several recommendations to prevent workplace and insurance discrimination. The highlights of these recommendations, which may be taken into account in developing legislation to prevent genetic discrimination, may be found at elsi/legislat.html ORNL.org.
  13. ^ Ibid
  14. ^ U.S. Department of Energy Human Genome Program, supra note 6
  15. ^ A number of scientists have called for the use the term “nuclear transplantation,” instead of “therapeutic cloning,” to help reduce public confusion. The term “cloning” has become synonymous with “somatic cell nuclear transfer,” a procedure that can be used for a variety of purposes, only one of which involves an intention to create a clone of an organism. They believe that the term “cloning” is best associated with the ultimate outcome or objective of the research and not the mechanism or technique used to achieve that objective. They argue that the goal of creating a nearly identical genetic copy of a human being is consistent with the term “human reproductive cloning,” but the goal of creating stem cells for regenerative medicine is not consistent with the term “therapeutic cloning.” The objective of the latter is to make tissue that is genetically compatible with that of the recipient, not to create a copy of the potential tissue recipient. Hence, “therapeutic cloning” is conceptually inaccurate. B. Vogelstein, B. Alberts, and K. Shine, “Please Don’t Call It Cloning!”, Science (15 February 2002), 1237
  16. ^ D. Cameron, “Stop the Cloning”, Technology Review, 23 May 2002’. Also available from Techreview.com, [hereafter “Cameron”]
  17. ^ M.C. Nussbaum and C.R. Sunstein, Clones And Clones: Facts And Fantasies About Human Cloning (New York: W.W. Norton & Co., 1998), 11. However, there is wide disagreement within scientific circles whether human cloning can be successfully carried out. For instance, Dr. Rudolf Jaenisch of Whitehead Institute for Biomedical Research believes that reproductive cloning shortcuts basic biological processes, thus making normal offspring impossible to produce. In normal fertilization, the egg and sperm go through a long process of maturation. Cloning shortcuts this process by trying to reprogram the nucleus of one whole genome in minutes or hours. This results in gross physical malformations to subtle neurological disturbances. Cameron, supra note 30
  18. ^ This op-ed appeared in the July 11, 1997, edition of The Washington Times, cartercenter.org
  19. ^ Asian Development Bank, Agricultural Biotechnology, Poverty Reduction and Food Security (Manila: Asian Development Bank, 2001). Also available from ADB.org
  20. ^ a b D. Bruce and A. Bruce, Engineering Genesis: The Ethics of Genetic Engineering, London: Earthscan Publications, 1999
  21. ^ S. Abdulla. “Drought Stress” Nature: Science Update; available from Nature.com, nsu; accessed 3 May 2002.
  22. ^ National Academy of Sciences. Transgenic Plants and World Agriculture (Washington: National Academy Press, 2001)
  23. ^ For an account of the research and development of Flavr Savr tomato, see B. Martineau, First Fruit: The Creation of the Flavr Savr Tomato and the Birth of Biotech Food (New York: McGraw-Hill, 2001)
  24. ^ A.F. Krattiger, An Overview of ISAAA from 1992 to 2000, ISAAA Brief No. 19-2000, 9
  25. ^ EuropaBio - An animal friendly alternative for cheeze makers, Europabio.org
  26. ^ EuropaBio - Biologically better bread, Europabio.org
  27. ^ L. P. Gianessi, C. S. Silvers, S. Sankula and J. E. Carpenter. Plant Biotechnology: Current and Potential Impact for Improving Pest management in US Agriculture, An Analysis of 40 Case Studies (Washington, D.C.: National Center for Food and Agricultural Policy, 2002), 5–6
  28. ^ C. James, “Global Review of Commercialized Transgenic Crops: 2002”, ISAAA Brief No. 27-2002, at 11–12. Also available from ISAAA.org
  29. ^ Pascual DW (2007). "Vaccines are for dinner". Proc Natl Acad Sci USA 104 (26): 10757–8. doi:10.1073/pnas.0704516104. PMID 17581867. http://www.pnas.org/cgi/content/full/104/26/10757. 
  30. ^ SemBioSys.ca
  31. ^ SemBioSys.ca
  32. ^ Monsanto and the Roundup Ready Controversy, - SourceWatch.org
  33. ^ Monsanto, - SourceWatch.org
  34. ^ Diaz E (editor). (2008). Microbial Biodegradation: Genomics and Molecular Biology (1st ed.). Caister Academic Press. ISBN 978-1-904455-17-2. ISBN 1904455174. http://www.horizonpress.com/biod. 
  35. ^ Martins VAP et al (2008). "Genomic Insights into Oil Biodegradation in Marine Systems". Microbial Biodegradation: Genomics and Molecular Biology. Caister Academic Press. ISBN 978-1-904455-17-2. http://www.horizonpress.com/biod. 
  36. ^ NIGMS.NIH.gov

Further reading

External links


Study guide

Up to date as of January 14, 2010

From Wikiversity

Biotechnology combines disciplines like genetics, molecular biology, biochemistry, embryology and cell biology, which are in turn linked to practical disciplines like chemical engineering, information technology, and robotics.

Biotechnology can also be defined as the manipulation of organisms to do practical things and to provide useful products.

One aspect of biotechnology is the directed use of organisms for the manufacture of organic products (examples include beer and milk products). For another example, naturally present bacteria are utilized by the mining industry in bioleaching. Biotechnology is also used to recycle, treat waste, clean up sites contaminated by industrial activities (bioremediation), and produce biological weapons.

There are also applications of biotechnology that do not use living organisms. Examples are DNA microarrays used in genetics and radioactive tracers used in medicine.

Red biotechnology is applied to medical processes. Some examples are the designing of organisms to produce antibiotics, and the engineering of genetic cures through genomic manipulation.

White biotechnology, also known as grey biotechnology, is biotechnology applied to industrial processes. An example is the designing of an organism to produce a useful chemical. White biotechnology tends to consume less in resources than traditional processes used to produce industrial goods.

Green biotechnology is biotechnology applied to agricultural processes. An example is the designing of transgenic plants to grow under specific environmental conditions or in the presence (or absence) of certain agricultural chemicals. One hope is that green biotechnology might produce more environmentally friendly solutions than traditional industrial agriculture. An example of this is the engineering of a plant to express a pesticide, thereby eliminating the need for external application of pesticides. An example of this would be Bt corn. Whether or not green biotechnology products such as this are ultimately more environmentally friendly is a topic of considerable debate.


Wikibooks

Up to date as of January 23, 2010

From Wikibooks, the open-content textbooks collection


This book is meant for students and professionals who are looking for reference on different areas in this field, to bring a new student or new hire up to speed.

A scientific revolution less than 20 years old that's already changing the foods we eat and react to the environment.

To bring out the best in nature.

Contents

What is Biotech?

"Defining "biotechnology""

  1. The use of living things to make products.
  2. The application of a biological process to produce a useful product.
  3. Any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific uses.

The use of microorganisms (such as bacteria or yeasts) or biological substances (such as enzymes) to perform specific industrial or manufacturing processes. Applications include the production of certain drugs, synthetic hormones, and bulk foodstuffs, as well as the bioconversion of organic waste and the cleanup of oil spills.

The application of the principles of engineering and technology to the life sciences--bioengineering.

Cloning, genetic manipulation, cell fusion, and mutation.

Essentially, doing "more and faster" what we have known and done for centuries. It is almost as old as agriculture iself.

Farmers and bakers are the biotechs of days gone by. Remember Grandma's freshly baked bread? How Grandpa kept the seeds of those really big pepper or tomatoes? Your grandparents were practicing biotechnology.

Modifying the genetic material of organisms directly and precisely. It enables the transfer of genes between diverse organisms, allowing combinations unlikely to occur by conventional means. Allowing speedier and more specific results.

Life- Defined as:

Moving
Growing
Eating
Breeding
Response to stimuli

Products

Products

Traditional

  1. Beer / Wine (See Fermentation)
  2. Yeast
  3. Ethanol
  4. Cheese

Protein Therapeutics

  1. Herceptin
  2. Neupogen
  3. Insulin - 1982 U.S. pharmaceutical manufacturer Eli Lilly the first to market genetically-engineered human insulin

Agriculture

  1. Bacillus thuringiensis (Bt) Crops Bt crops
  2. Herbicide tolerance
  3. Disease resistance
  4. Pest resistance

Industrial / Environmental

  1. Hazardous Waste
  2. Wastewater Treatment
  3. Bioremediation

Other

  1. Plastics
  2. Biopulping
  3. GMO (Genetically modified organism)

21 CFR 58

Good laboratory practice for nonclinical laboratory studies:

http://www.access.gpo.gov/nara/cfr/waisidx_02/21cfr58_02.html

21 CFR 11

Title 21 Code of Federal Regulations (21 CFR Part 11) Electronic Records; Electronic Signatures

http://www.fda.gov/ora/compliance_ref/part11/

21 Code of Federal Regulations Parts 210 and 211

Part 210 - current good manufacturing practice in manufacturing, processing, packing, or holding of drugs; general

Part 211 - current good manufacturing practice for finished pharmaceuticals

http://www.fda.gov/cder/dmpq/cgmpregs.htm

SOPs

SOP's (Standard Operating Procedures)

  1. Approved by management
  2. You don't put data in a SOP
  3. Written at 6th grade level
  4. Clear, current, complete
  5. Simple, Concise, Accurate
  6. Archived Where?

Notebooks

Notebook

  1. If it is not written, it is not done.
  2. pages signed
  3. Can you read it?
  4. Black or blue ink
  5. pages numbered
  6. Leave some pages in the front for TOC
  7. Required by GLP / GMP regs
  8. Signed by witness
  9. Traceability

Documentation for Integrity and traceablity

  1. What you did
  2. Why you did it
  3. Results
  4. Reference earlier work?
  5. Also for Quality

Keys to Successful Biotech products

Keys to Successful Biotech products

  1. Availability of labor force
  2. Skilled labor force
  3. Power
  4. Good Universities nearby:
   * For retraining
   * For new ideas
  1. National and international labor pool
  2. Raw materials
  3. Marketing
   * Clear trend and need
  1. FDA approval

Record Keeping

   Inventory control logs
   Process Steps
   SOP
   et cetera
   If it's not written it's not done
       Love it
       Live it
   * Clinical trials
   * GMP apply to:
               + Manufacture
               + Processing
               + Packing
               + Storage
   * Validation of each process
   * PAT (Process Analytical Technology)
     Quality should be built in and by design
  1. Environmental Concerns
   * Use of energy
   * Use of water
   * Environmental Laws
   * Waste streams
  1. Do we have the technology?

requirements

   Safety
   Identity and Strength
   Quality and purity
    The plant:
          Can we Measure
               + pH
               + Aeration
               + Baffiling
               + impellers
               + medium
               + Seed train
  1. Are we Better than our competition?
why?

processes

Development / Upstream / Downstream processes

Development

  1. Need / market analysis
  2. Select a solution
  3. Process Development
  4. Understand the growth requirements
  5. Calculate yields of biomass
  6. Run small scale test

Chemical

Yeast

 High biomass yields
 Will secrete the protein
 Picha Pastoris

Fungi

Mammalian Cells

Upstream

  1. Media prep

Fermentation?

Find best conditions

Expensive Labor intensive Open Ended Time Consuming

Constraints

Raw Materials Batch to Batch variations Transportation costs Storage

  1. Seed Prep

Find cell line

Composition Growth kinetics Yield Seed Bank

Master Seed Bank MSB

Original Stored Cells

Working Seed Bank WSB

Used in actual fermentation

Downstream

  1. Slurry Handling
  2. Excreted product
  3. Contamination
  4. Product purification
  5. Separation / purification / sterilization
How?
  1. Plate and frame
  2. Bulk filtration
  3. Spiral wound
       Plugs up easily
       Usually not used for bulk filtration
  1. Storage

Technician: Skills Needed

The Biotech Technician must be a person possessing skills with ability to solve problems and meet the customer in such a way that the translations of what is possible can be made clear. They have to maintain a notebook, one that can be read by someone else. Present results in a clear manner, and work with others to meet objectives.

Laboratory Skills

A technician must use the tools of the trade not unlike any other trade, we are farmers but our herd is tiny tiny wildlife. To take care of our herd we must measure certain aspects of their environment.

Solute,solvent, and solution

Solute = Dry Material
Solvent = What you mix with
Solution = Solute + Solvent

pH

Measurement
  1. Probe and meter

most accurate more expensive piece of equipment Store in buffer Check for clogging

  1. Litmus paper

very coarse measurement of pH

  1. Field kit

The letters pH stand for "power of hydrogen"

Hydrogen the most abundant element in the universe is hydrogen, which makes up about 3/4 of all matter!

Stronger acids give up more protons, H+ (hydrogen ions); stronger bases give up more OH- (hydroxide ions). Neutral substances have an even balance of H+ and OH-, Eg. Pure (distilled) water.

>7 base -- 7 Neutral -- <7 Acid

Depending on your definition, an acid is a hydrogen ion or proton donator and a base is a hydrogen ion acceptor, hydroxide ion donator, or electron acceptor.

Acids produce H+ ions in aqueous solutions, whereas bases produce OH- ions in aqueous solutions

pH electrode compared to a battery

Store in buffer not H2O

Mercury tube Good for metals and biologicals and up to 80 degrees C

The common Silver-Silver Chloride reference electrode used with most combination pH electrodes has a Potassium Chloride salt-bridge which is saturated with Silver Chloride.

Works well in most samples, but not in biological samples containing proteins or related materials

Calibration

Span error Difference b/w perfect and actual pH Electrode at 25C produces 59.12 mV/pH unit

Offset error

Difference from Perfect reading from actual reading is the offset error

signal @ pH 7.0 @ 25 C is 0 mV

Three point calibration

pH's 4, 7 and 10

Calibrate W/I range you going to use

Buffer and reagent Prep

Chemist use buffers to moderate the pH of a reaction. Buffers stabilize a solution at a specific pH value. Resist pH change when small amounts of acid or alkali are added.

KPO4

KPO4 buffer is highly recommended for most P450 assays (microsomal or recombinant enzymes) with the exception of CYP 2C9 and and 2A6 where a Tris buffer system is more appropriate.

TRIS buffer

TRIS buffers are used by biochemists to control pH in the physiological range (about 7 to 8 pH) because phosphates cause undesirable side reactions with the biological substances in their test samples.


"Good" buffers

These buffers were well received by the research community because "Good" buffers are nontoxic, easy to purify and their pKa is typically between 6.0 and 8.0, the range at which most biological reactions occur.

The "Good" buffers also feature minimal penetration of membranes, minimal absorbance in the 240-700 nm range and minimal effects due to salt, concentration or temperature.


pKa = dissociation constant


In chemistry and biochemistry, a dissociation constant or an ionization constant is a specific type of equilibrium constant used for dissociation (ionization) reactions. Dissociation in chemistry and biochemistry is a general process in which complexes, molecules, or salts separate or split into smaller molecules, ions, or radicals, usually in a reversible manner. Dissociation is the opposite of association and recombination.

Problems

  1. Ionic Strength
  2. Temperature
  3. Counter ion

colony agar plating

What is Agar?

A gelatinous material derived from certain marine algae.

  1. Growth Media
  2. Carbon Sources
  3. Carbohydrates
  4. Proteins
  5. Nitrogen Sources

Two types:

Simple
Complex

Components required for preparing a minimal agar

  1. minerals
  2. H2O
  3. vitamins
  4. Precursors
  5. Compounds that may lead to your product - inducers
  6. Compounds added to induce gene expression

LB (Luria-Bertani) Media

Agar Plates
Blood

contains blood cells from an animal (e.g. a sheep). Most bacteria will grow on this medium

Chocolate

This contains lysed blood cells, and is used for growing fastidious (fussy) respiratory bacteria.

Mannitol Salt

Purpose Mannitol salt agar is both a selective and differential growth medium.

Brilliant Green

Inhibits Gram+ MacConkey

This type of agar is used since it is one of the most forgiving media available - it is hard to contaminate, and E. coli usually grow up as red colonies.

(Almost all spore forming bacteria are Gram-positive, but these cannot grow on MacConkey agar because of the detergent in it (bile salts), and very few Gram-negative bacteria can tolerate either the initial dryness of the plates, or the boiling temperatures needed to make the MacConkey agar. Also, while fungal spores can tolerate the dryness, they cannot tolerate the boiling.)

This is an agar upon which only Gram-negative bacteria can grow

Starch

An agar plate is a sterile Petri dish that contains agar plus nutrients, and is used to culture bacteria or fungi.

Neomycin agar

contains the antibiotic neomycin.

Sabouraud agar

Used for fungi. It contains gentamicin and has a low pH that will kill most bacteria.

LB

+ Complex + pH 7.2

UV/VIS Spectroscopy

Common UV/ VIS spectrophotometers Following is a list of commonly used spectrophotometers: GeneSys 20 HP8452A Diode Array Spectronic 20

Ultraviolet-Visible spectroscopy or Ultraviolet-Visible spectrophotometry (UV/ VIS) involves the spectroscopy of photons (spectrophotometry). It uses light in the visible and adjacent near ultraviolet (UV) and near infrared (NIR) ranges. In this region of energy space molecules undergo electronic transitions.

A=elc

  1. A=Absorbance
  2. e=Molar Absorbtivity
  3. l=Path length (cm)
  4. c=Concentration (M)

Laboratory calculations

  1. + - Prep of Sterile solid and Liquid media

Sterilization Methods

There are different types of Sterilization techniques. Some of them are 1. Physical sterilization 2. Chemical sterilization

Under Physical sterilization a) Heat b) Filtration c) Ionising Radiation etc., In Heat sterilization i. Temperature above 100 C ii. Temperature at 100 C iii. Temperature below 100 C.

i. Temperature above 100 C There are two methods involved in it a. Moisture heat sterilization b. Dry heat sterilization

Pipetting

  1. Growing concerns regarding repetitive strain injuries (RSIs)
  2. Adopting good pipetting techniques is critical for the accuracy of your analysis and your health.
  3. Contamination Prevention
  4. Care of

Measuring / Mixing

Using a balance Calibration / documentation

UV/VIS spectroscopy

   * http://www.scienceofspectroscopy.info/
   * Use correct Reagent
   * Use correct Wavelength

Gel Electrophoresis

Gel electrophoresis is a method that separates macromolecules-either nucleic acids or proteins-on the basis of size, electric charge, and other physical properties.

materials

Two basic types
agarose and polyacrylamide

agarose

Agarose is a natural colloid extracted from sea weed It is very fragile and easily destroyed by handling Agarose gels have very large "pore" size and are used primarily to separate very large molecules with a molecular mass greater than 200 kDaltons Agarose gels can be processed faster than polyacrylamide gels, but their resolution is inferior.

Agarose is a linear polysaccharide (average molecular mas about 12,000) made up of the basic repeat unit agarobiose, which comprises alternating units of galactose and 3,6-anhydrogalactose. Agarose is usually used at concentrations between 1% and 3%. Agarose is a chain of sugar molecules, and is extracted from seaweed.

Perhaps you have seen the terms TBE or TAE.

These are names of two commonly used buffers in electrophoresis.

The "T" stands for Tris, a chemical which helps maintain a consistent pH of the solution.

The "E" stands for EDTA, which itself is another anacronym. EDTA chelates (gobbles up) divalent cations like magnesium. This is important because most nucleases require divalent cations for activity, and you certainly wouldn't want any stray nucleases degrading your sample while it's running through the gel, would you?

Finally, the "B" or "A" stand for Boric acid or Acetic acid, which provide the proper ion concentration for the buffer.

polyacrylamide

The polyacrylamide gel electrophoresis (PAGE) technique was introduced by Raymond and Weintraub (1959).

Polyacrylamide is the same material that is used for skin electrodes and in soft contact lenses.

provide a wide variety of electrophoretic conditions:

By controlling the percentage (from 3% to 30%), precise pore sizes can be obtained, usually from 5 to 2,000 kdal. Polyacrylamide gels can be cast in a single percentage or with varying gradients Polyacrylamide gels offer greater flexibility and more sharply defined banding than agarose gels.

Spectrophotometer

Absorbance
Transmission
Colormetric
400 to 700 nm
Pertaining to measurement of concentration of a solution based on its absorption or transmission of light, or on the intensity of color in a liquid.

Centrifugation

  • Speed
  • Distance from Center of rotation
  • RCF= relative Centrifugal Force

factors

         o Higher RCF the faster the sedimentation
         o viscosity
         o size of particle
         o difference b/w particle and medium

Application

         o Seperation
         o + - Safety
               + Never exceed Max speed of rotor
               + Never use a cracked tube or bottle
         o + - Maintenance
               + Clean with mild detergent
               + air dry rotor
               + Check rotor O-rings

Types

+ - low speed

               + rpm<10k
               + g<8000

+ - High Speed

               + rpm<30k
               + g<100k
               + refrigeration used

+ - ultracentrifuge

               + rpm<120k
               + g<700k
               + refrigeration used
               + vacuum

+ - microfuge

               + tabletop
               + rpm<15k
               + g<21k
               + 1-2ml volumes

parts

               + + - rotors
                     # horizontal
                     # + - fixed angle
                           * b/w 15 & 40 degrees
                     # vertical

tubes / bottles

                     # glass
                     # stainless steel
                     # polycarbonate
                     # + - Teflon
                           * expensive
                     # + - polypropylene
                           * What most people use

Aseptic techniques

Aseptic techniques is defined as a method that keeps undesirable microbes from contaminating a pure culture.

Only a single species of an organism is what one should work with in a microorganismal laboratory all materials that will be used for transfer, growth, and experimentation of the microbe must be sterilized media and glassware most often is sterilized by using an autoclave.

Lab bench needs to be clean with a disinfectant before and after working.

Hands should be washed upon entering and leaving the laboratory.

Inoculation tools and the tops of tubes need to be sterilized over an flame

Inoculating Loop - held like a pencil

fermenter design & application

Employee traits

Computer Skills

  1. Popular OS'es and file management
  2. Excel
  3. PowerPoint
  4. Word
  5. Linux

People Skills

Energy and interest

WHAT CAN I DO FOR YOU?

How you are qualified

Desire to continue learning

Brainpower:

In today's world, it's "be sharp or die."

Because we are status quo creatures. We like things just the way they are, thank you. Change unsettles us. If it works (or appears to work, actually), don't fix it.

Ongoing education / cross pollination of other areas:

Basic Microbiology

Basic Microbiology

Cell Identification

Enterotube

cell growth

Log Phase

cell maintenance & storage

Cryopreservation

http://nalgenelab.nalgenunc.com/techdata/technical/manual.asp

The protection of biological molecules during freezing and freeze-drying( lyophilization) is a subject of considerable practical importance, particularly in the pharmaceutical industry.
   wide variety of compounds as cryoprotectants
Saccharides are often used in this capacity
They have been found to protect proteins during freezing and drying stresses.
They have also been shown to prevent damage to cells during freezing and drying.

Basic Molecular Biology

Basic Molecular Biology

Organism Design

Genetic Analysis

Strain Validation Applications

Genetic engineering

"Geneticist and science writer Steve Jones argues that humanity does not, and will never have the technology that proponents of transhumanism seek. He once joked that the letters of the genetic code, A, C, G and T should be replaced with the letters H, Y, P and E. Jones claims that technologies like genetic engineering will never be as powerful as is popularly believed."

-http://en.wikipedia.org/wiki/Transhumanism

E. Coli The workhorse of molecular biology

Theodor Escherich isolates a microbe from the colon that is later given the name Escherichia coli in his honor.

  1. Ecoli
    1. HB101
      1. Fairly safe
      2. Needs
      3. 37 C
      4. Food
      5. H2O
      6. Vitamins

Escherichia coli, a subgroup of fecal coliform bacteria that is present in the intestinal tracts and feces of warm-blooded animals.

It is used as an indicator of the potential presence of pathogens. There are many different strains of E. coli that are classified into more than 170 serogroups.

Although most strains of E. coli are harmless and live in the intestines of healthy humans and animals, the E. coli O157:H7 strain produces a powerful toxin and can cause severe illness.

Its presence in groundwater is a common indicator of fecal contamination.

("Enteric" is the adjective that describes organisms that live in the intestines. "Fecal" is the adjective for organisms that live in feces, so it is often a synonym for "enteric.") The name comes from its discoverer, Theodor Escherich.

Basic Protein Separation

Basic Protein Separation

  1. Protein Separation
  2. Protein Analysis

Basic Tissue Culture

Basic Tissue Culture

  1. Cultures
  2. Cell line
  3. Culture methods
  4. Animal handling

Basic Chromatography

Basic Chromatography

  1. HPLC Systems

High Performance Liquid Chromatography (HPLC):

HPLC is a popular method of analysis because it is easy to learn and use and is not limited by the volatility or stability of the sample compound

Modern HPLC has many applications including separation, identification, purification, and quantification of various compounds

Textbook on High Performance Liquid Chromatography (HPLC) http://hplc.chem.shu.edu/NEW/HPLC_Book/

GC-MS

Biotech HOT SPOTS in the US

  1. RTP - Research Triangle Park, created in 1959, located between Duke University in Durham, North Carolina, the State University in Raleigh, and the University of North Carolina at Chapel Hill.
  2. Boston
  3. Albany
  4. South San Francisco

Acronyms / definitions

Acronyms / definitions

GLP

GLP Good Laboratory Practices

GMP

Good Manufacturing Practice http://www.fda.gov/cder/dmpq/cgmpregs.htm

Biocidol

   Kills cells
   Disinfectant

Biostat

   Chemical that stops cell growth - doesnt kill

Supernatants

Liquid removed from a tank once the solids have settled. Usually a clear liquid left after material (like cells) has been precipitated or centrifuged.

The material remaining above the pellet after centrifugation of a suspension.

Authors of this Wikibook

This Wikibook was written by Tom Maioli tmaioli


Simple English

Biotechnology is a technology or science based on biology, especially when this is used in agriculture, food science, and medicine.

Biotechnology is often used to refer to genetic engineering technology of the 21st century, however the term encompasses a wider range and history of procedures for modifying biological organisms according to the needs of humanity, going back to the initial modifications of native plants into improved food crops through artificial selection and hybridization. Bioengineering is the science upon which all Biotechnological applications are based. With the development of new approaches and modern techniques, traditional biotechnology industries are also acquiring new horizons enabling them to improve the quality of their products and increase the productivity of their systems.

Biotechnology has also made cloning (the process duplicating organisms) possible, a lot of people think that this is morally wrong while others think it could solve many diseases.

Biotechnology can be used to solve a great number of problems, ranging from product efficiency to reducing global warming.rue:Біотехнолоґія








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