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DNA technology and forensics
DNA is used for identification in forensic science.
DNA can be used to identify any type of living organism. This is done by analyzing DNA
sequences that are unique to a species. The field of forensic science uses DNA analysis toidentify individuals. While it is not yet possible to identify individuals by exact DNA matches, it
is possible to analyze DNA regions that give a very high probability for matching the individual
with the sample.
What is GMO?Agricultural Crops That Have a Risk of Being GMO
GMOs, or genetically modified organisms, are plants or animals created through the genesplicing techniques of biotechnology (also called genetic engineering, or GE). This experimental
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technology merges DNA from different species, creating unstable combinations of plant, animal,
bacterial and viral genes that cannot occur in nature or in traditional crossbreeding.
For consumers, it can be difficult to stay up-to-date on food ingredients that are at-risk of being
genetically modified, as the list of at-risk agricultural ingredients is frequently changing. As part
of the Non-GMO Projects commitment to informed consumer choice, we work diligently tomaintain an accurate list of risk ingredients.
Agricultural products are segmented into two groups: (1) those that are high-riskof being GMObecause they are currently in commercial production, and (2) those that have a monitored risk
because suspected or known incidents of contamination have occurred and/or the crops have
genetically modified relatives in commercial production with which cross-pollination (andconsequently contamination) is possible. For more information on the Non-GMO Projects
testing and verification of risk ingredients and processed foods, please see the Non-GMO Project
Standard.
High-Risk Crops (in commercial production; ingredients derived from these must be testedevery time prior to use in Non-GMO Project Verified products (as of December 2011):
Alfalfa (first planting 2011)
Canola (approx. 90% of U.S. crop)
Corn (approx. 88% of U.S. crop in 2011)
Cotton (approx. 90% of U.S. crop in 2011)
Papaya (most of Hawaiian crop; approximately 988 acres)
Soy (approx. 94% of U.S. crop in 2011)
Sugar Beets (approx. 95% of U.S. crop in 2010)
Zucchini and Yellow Summer Squash (approx. 25,000 acres)
ALSO high-risk: animal products (milk, meat, eggs, honey, etc.) because of contamination in
feed.
Monitored Crops (those for which suspected or known incidents of contamination have
occurred, and those crops which have genetically modified relatives in commercial productionwith which cross-pollination is possible; we test regularly to assess risk, and move to High-
Risk category for ongoing testing if we see contamination):
Beta vulgaris (e.g., chard, table beets)
Brassica napa (e.g., rutabaga, Siberian kale)
Brassica rapa (e.g., bok choy, mizuna, Chinese cabbage, turnip, rapini, tatsoi)
Curcubita (acorn squash, delicata squash, patty pan) Flax
Rice
Common Ingredients Derived from GMO Risk CropsAmino Acids, Aspartame, Ascorbic Acid, Sodium Ascorbate, Vitamin C, Citric Acid, Sodium
Citrate, Ethanol, Flavorings (natural and artificial), High-Fructose Corn Syrup, Hydrolyzed
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Vegetable Protein, Lactic Acid, Maltodextrins, Molasses, Monosodium Glutamate, Sucrose,
Textured Vegetable Protein (TVP), Xanthan Gum, Vitamins, Yeast Products.
You may also be wondering about
Tomatoes: In 1994, genetically modified Flavr Savr tomatoes became the first commerciallyproduced GMOs. They were brought out of production just a few years later, in 1997, due to
problems with flavor and ability to hold up in shipping. There are no genetically engineered
tomatoes in commercial production, and tomatoes are considered low-risk by the Non-GMO
Project Standard.
Potatoes: Genetically modified NewLeaf potatoes were introduced by Monsanto in 1996. Due to
consumer rejection several fast-food chains and chip makers, the product was never successful
and was discontinued in the spring of 2001. There are no genetically engineered potatoes in
commercial production, and potatoes are considered low-risk by the Non-GMO Project
Standard.
Wheat: There is not currently, nor has there ever been, any genetically engineered wheat on the
market. Ofall low-risk crops, this is the one most commonly (and incorrectly) assumed to be
GMO. It is a key commodity crop, and the biotech industry is pushing hard to bring GMO
varieties to market. The Non-GMO Project closely watches all development on this front.
Salmon: A company called AquaBounty is currently petitioning the FDA to approve its
genetically engineered variety of salmon, which has met with fierce consumer resistance.Find
out more here.
Pigs: A genetically engineered variety of pig, called Enviropig was developed by scientists at the
University of Guelph, with research starting in 1995 and government approval sought beginning
in 2009. In 2012 the University announced an end to the Enviropig program, and the pigs
themselves were euthanized in June 2012.
DNA Technology Applications
The use ofrecombinant DNA technology has become commonplace as new products fromgenetically altered plants, animals, and microbes have become available for human use. In 1997,
Dolly made headlines as the first successfully clonedlarge mammal (sheep). Since then therehave been many similar advances in medicine, such as treatments for cancer; many advances in
agriculture, such as transgenic insect-resistant crops; and many advances in animal husbandry,
such as growth hormones and transgenic animals (an animal that has received recombinant
DNA).
Most biotechnologists envision DNA technological applications as one of the new frontiers inscience with tremendous growth and discovery potential.
Medicine
Genetic engineering has resulted in a series of medical products. The first two commercially
prepared products from recombinant DNA technology were insulin and human growth hormone,
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both of which were cultured in the E. coli bacteria. Since then a plethora of products have
appeared on the market, including the following abbreviated list, all made in E. coli:
Bionote
A vaccine is usually a harmless version of a bacterium or virus that is injected into an organismto activate the immune system to attack and destroy similar substances in the future.
Tumor necrosis factor. Treatment for certain tumor cells
Interleukin-2 (IL-2). Cancer treatment, immune deficiency, and HIV infection treatment
Prourokinase. Treatment for heart attacks
Taxol. Treatment for ovarian cancer
Interferon. Treatment for cancer and viral infections
In addition, a number ofvaccines are now commercially prepared from recombinant hosts. At
one time vaccines were made by denaturing the disease and then injecting it into humans with
the hope that it would activate their immune system to fight future intrusions by that invader.Unfortunately, the patient sometimes still ended up with the disease.
With DNA technology, only the identifiable outside shell of the microorganism is needed,copied, and injected into a harmless host to create the vaccine. This method is likely to be much
safer because the actual disease-causing microbe is not transferred to the host. The immune
system is activated by specific proteins on the surface of the microorganism -e. DNA technologytakes that into account and only utilizes identifying surface features for the vaccine. Currently
vaccines for the hepatitis B virus, herpes type 2 viruses, and malaria are in development for trial
use in the near future.
Agriculture
Crop plants have been and continue to be the focus of biotechnology as efforts are made toimprove yield and profitability by improving crop resistance to insects and certain herbicides and
delaying ripening (for better transport and spoilage resistance). The creation of a transgenic
plant, one that has received genes from another organism, proved more difficult than animals.Unlike animals, finding a vector for plants proved to be difficult until the isolation of the Ti
plasmid, harvested from a tumor-inducing (Ti) bacteria found in the soil. The plasmid is shot
into a cell, where the plasmid readily attaches to the plant's DNA. Although successful in fruitsand vegetables, the Ti plasmid has generated limited success in grain crops.
Creating a crop that is resistant to a specific herbicide proved to be a success because theherbicide eliminated weed competition from the crop plant. Researchers discovered herbicide-
resistant bacteria, isolated the genes responsible for the condition, and shot them into a crop
plant, which then proved to be resistant to that herbicide. Similarly, insect-resistant plants arebecoming available as researchers discover bacterial enzymes that destroy or immobilize
unwanted herbivores, and others that increase nitrogen fixation in the soil for use by plants.
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Geneticists are on the threshold of a major agricultural breakthrough. All plants need nitrogen to
grow. In fact, nitrogen is one of the three most important nutrients a plant requires. Although the
atmosphere is approximately 78 percent nitrogen, it is in a form that is unusable to plants.However, a naturally occurring rhizobium bacterium is found in the soil and converts
atmospheric nitrogen into a form usable by plants. These nitrogen-fixing bacteria are also found
naturally occurring in the legumes of certain plants such as soybeans and peanuts. Because theycontain these unusual bacteria, they can grow in nitrogen-deficient soil that prohibits the growthof other crop plants. Researchers hope that by isolating these bacteria, they can identify the DNA
segment that codes for nitrogen fixation, remove the segment, and insert it into the DNA of a
profitable cash crop! In so doing, the new transgenic crop plants could live in new fringeterritories, which are areas normally not suitable for their growth, and grow in current locations
without the addition of costly fertilizers!
Animal Husbandry
Neither the use of animal vaccines nor adding bovine growth hormones to cows to dramatically
increase milk production can match the real excitement in animal husbandry: transgenic animalsand clones.
Transgenic animals model advancements in DNA technology in their development. The
mechanism for creating one can be described in three steps:
1. Healthy egg cells are removed from a female of the host animal and fertilized in the laboratory.
2. The desired gene from another species is identified, isolated, and cloned.
3. The cloned genes are injected directly into the eggs, which are then surgically implanted in the
host female, where the embryo undergoes a normal development process.
It is hoped that this process will provide a cheap and rapid means of generating desired enzymes,other proteins, and increased production of meat, wool, and other animal products throughcommon, natural functions.
Ever since 1997 when Dolly was cloned, research and experimentation to clone useful livestockhas continued unceasingly. The attractiveness of cloning is the knowledge that the offspring will
be genetically identical to the parent as in asexual reproduction. Four steps describe the general
process:
1. A differentiated cell, one that has become specialized during development, with its diploid
nucleus is removed from an animal to provide the DNA source for the clone.
2. An egg cell from a similar animal is recovered and the nucleus is removed, leaving only the
cytoplasm and cytoplasm organelles.
3. The two egg cells are fused with an electric current to form a single diploid cell, which then
begins normal cell division.
4. The developing embryo is placed in a surrogate mother, who then undergoes a normal
pregnancy.
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Completed in 2003, the Human Genome Project (HGP) was a 13-year projectcoordinated by the U.S. Department of Energy and the National Institutes of Health.
During the early years of the HGP, the Wellcome Trust (U.K.) became a major partner;
additional contributions came from Japan, France, Germany, China, and others. See
ourhistory page for more information.
Project goals were to
identifyall the approximately 20,000-25,000 genes in human DNA,
determine the sequences of the 3 billion chemical base pairs that make uphuman DNA,
store this information in databases,
improve tools for data analysis, transfer related technologies to the private sector, and address the ethical, legal, and social issues (ELSI) that may arise from the
project.
Though the HGP is finished, analyses of the data will continue for many years. Follow
this ongoing research on ourMilestones page. An important feature of the HGP project
was the federal government's long-standing dedication to the transfer of technology to
the private sector. By licensing technologies to private companies and awarding grants
for innovative research, the project catalyzed the multibillion-dollar U.S. biotechnology
industry and fostered the development of new medical applications.
To help achieve these goals, researchers also studied the genetic makeup of several nonhuman
organisms. These include the common human gut bacteriumEscherichia coli, the fruit fly, and
the laboratory mouse.
A unique aspect of the U.S. Human Genome Project is that it was the first large scientificundertaking to address potential ELSI implications arising from project data.
Another important feature of the project was the federal government's long-standing dedication
to the transfer of technology to the private sector. By licensing technologies to private companies
and awarding grants for innovative research, the project catalyzed the multibillion-dollar U.S.biotechnology industry and fostered the development of new medical applications.
Landmark papers detailing sequence and analysis of the human genome were published in
February 2001 and April 2003 issues ofNature and Science. See an index of these papers and
learn more about the insights gained from them
http://www.ornl.gov/sci/techresources/Human_Genome/project/hgp.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/hg5yp/index.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/progress.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/privatesector.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/privatesector.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/medicine/medicine.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/journals/journals.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/journals/insights.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/journals/insights.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/journals/journals.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/medicine/medicine.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/privatesector.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/privatesector.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/progress.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/hg5yp/index.shtmlhttp://www.ornl.gov/sci/techresources/Human_Genome/project/hgp.shtml7/29/2019 DNA Technology and Forensics
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What is gene therapy?
Gene therapy is an experimental technique that uses genes to treat or prevent disease. In the
future, this technique may allow doctors to treat a disorder by inserting a gene into a patients
cells instead of using drugs or surgery. Researchers are testing several approaches to genetherapy, including:
Replacing a mutated gene that causes disease with a healthy copy of the gene.
Inactivating, or knocking out, a mutated gene that is functioning improperly.
Introducing a new gene into the body to help fight a disease.
Although gene therapy is a promising treatment option for a number of diseases (including
inherited disorders, some types of cancer, and certain viral infections), the technique remains
risky and is still under study to make sure that it will be safe and effective. Gene therapy iscurrently only being tested for the treatment of diseases that have no other cures.
Biofertilizer
From Wikipedia, the free encyclopediaJump to: navigation, search
Tolypothrix, Cyanobacteria often used as fertilizer.
Blue-green algae cultured in specific media. Blue-green algae can be helpful in agriculture as
they have the -green algae is used as a bio-fertilizer.
http://en.wikipedia.org/wiki/Biofertilizer#mw-headhttp://en.wikipedia.org/wiki/Biofertilizer#p-searchhttp://en.wikipedia.org/wiki/Algaehttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Blue-green_algae_cultured_in_specific_media.jpghttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/File:Tolypothrix_(Cyanobacteria).JPGhttp://en.wikipedia.org/wiki/Algaehttp://en.wikipedia.org/wiki/Biofertilizer#p-searchhttp://en.wikipedia.org/wiki/Biofertilizer#mw-head7/29/2019 DNA Technology and Forensics
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A biofertilizer (also bio-fertilizer) is a substance which contains living microorganisms which,
when applied to seed, plant surfaces, or soil, colonizes the rhizosphere or the interior of the plant
and promotes growth by increasing the supply or availability of primary nutrients to the hostplant.
[1]Bio-fertilizers add nutrients through the natural processes ofnitrogen fixation,
solubilizing phosphorus, and stimulating plant growth through the synthesis of growth-
promoting substances. Bio-fertilizers can be expected to reduce the use ofchemical fertilizersand pesticides. The microorganisms in bio-fertilizers restore the soil's natural nutrient cycle andbuild soil organic matter. Through the use of bio-fertilizers, healthy plants can be grown, while
enhancing the sustainability and the health of the soil. Since they play several roles, a preferred
scientific term for such beneficial bacteria is "plant-growth promoting rhizobacteria" (PGPR).Therefore, they are extremely advantageous in enriching soil fertility and fulfilling plant nutrient
requirements by supplying the organic nutrients through microorganism and their byproducts.
Hence, bio-fertilizers do not contain any chemicals which are harmful to the living soil.
Bio-fertilizers eco friendly organic agro-input and more cost-effective than chemical fertilizers.
Bio-fertilizers such as Rhizobium, Azotobacter, Azospirillum and blue green algae (BGA) have
been in use a long time. Rhizobiuminoculant is used for leguminous crops. Azotobactercan beused with crops like wheat, maize, mustard, cotton, potato and other vegetable crops.
Azospirillum inoculations are recommended mainly forsorghum, millets, maize, sugarcane andwheat. Blue green algaebelonging to a general cyanobacteria genus,NostocorAnabaenaorTolypothrixorAulosira, fix atmospheric nitrogen and are used as inoculations for paddy crop
grown both under upland and low-land conditions.Anabaenain association with water fern
Azolla contributes nitrogen up to 60 kg/ha/season and also enriches soils with organic matter.[2]
Other types of bacteria, so-called phosphate-solubilizing bacteria, such as Pantoea agglomerans
strain P5 orPseudomonas putida strain P13,[3]
are able to solubilize the insoluble phosphate fromorganic and inorganic phosphate sources.
[4]In fact, due to immobilization of phosphate by
mineral ions such as Fe, Al and Ca ororganic acids, the rate of available phosphate (Pi) in soil is
well below plant needs. In addition, chemical Pi fertilizers are also immobilized in the soil,
immediately, so that less than 20 percent of added fertilizer is absorbed by plants. Therefore,reduction in Pi resources, on one hand, and environmental pollutions resulting from both
production and applications of chemical Pi fertilizer, on the other hand, have already demanded
the use of new generation of phosphate fertilizers globally known as phosphate-solubilizingbacteria or phosphate bio-fertilizers
Recombinant DNA (rDNA) molecules are DNA sequences that result from the use of
laboratory methods (molecular cloning) to bring together genetic material from multiple sources,creating sequences that would not otherwise be found in biological organisms. Recombinant
DNA is possible because DNA molecules from all organisms share the same chemical structure;
they differ only in the sequence ofnucleotides within that identical overall structure.
Consequently, when DNA from a foreign source is linked to host sequences that can drive DNAreplication and then introduced into a host organism, the foreign DNA is replicated along with
the host DNA.
http://en.wikipedia.org/wiki/Microorganismhttp://en.wikipedia.org/wiki/Rhizospherehttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-vessey2003-1http://en.wikipedia.org/wiki/Biofertilizer#cite_note-vessey2003-1http://en.wikipedia.org/wiki/Biofertilizer#cite_note-vessey2003-1http://en.wikipedia.org/wiki/Nitrogen_fixationhttp://en.wikipedia.org/wiki/Phosphorushttp://en.wikipedia.org/wiki/Chemical_fertilizershttp://en.wikipedia.org/wiki/Pesticideshttp://en.wikipedia.org/wiki/Plant-growth_promoting_rhizobacteriahttp://en.wikipedia.org/wiki/Soil_fertilityhttp://en.wikipedia.org/wiki/Chemical_fertilizershttp://en.wikipedia.org/wiki/Rhizobiumhttp://en.wikipedia.org/wiki/Azotobacterhttp://en.wikipedia.org/wiki/Blue_green_algaehttp://en.wikipedia.org/wiki/Rhizobiumhttp://en.wikipedia.org/wiki/Azotobacterhttp://en.wikipedia.org/wiki/Wheathttp://en.wikipedia.org/wiki/Maizehttp://en.wikipedia.org/wiki/Mustard_planthttp://en.wikipedia.org/wiki/Cottonhttp://en.wikipedia.org/w/index.php?title=Azospirillum&action=edit&redlink=1http://en.wikipedia.org/wiki/Sorghumhttp://en.wikipedia.org/wiki/Milletshttp://en.wikipedia.org/wiki/Maizehttp://en.wikipedia.org/wiki/Sugarcanehttp://en.wikipedia.org/wiki/Blue_green_algaehttp://en.wikipedia.org/wiki/Cyanobacteriahttp://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Nostochttp://en.wikipedia.org/wiki/Nostochttp://en.wikipedia.org/wiki/Nostochttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/w/index.php?title=Tolypothrix&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Tolypothrix&action=edit&redlink=1http://en.wikipedia.org/wiki/Aulosirahttp://en.wikipedia.org/wiki/Aulosirahttp://en.wikipedia.org/wiki/Aulosirahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Azollahttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-2http://en.wikipedia.org/wiki/Biofertilizer#cite_note-2http://en.wikipedia.org/wiki/Biofertilizer#cite_note-2http://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/Pantoea_agglomeranshttp://en.wikipedia.org/wiki/Pseudomonas_putidahttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-3http://en.wikipedia.org/wiki/Biofertilizer#cite_note-3http://en.wikipedia.org/wiki/Biofertilizer#cite_note-3http://en.wikipedia.org/wiki/Inorganic_phosphatehttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-4http://en.wikipedia.org/wiki/Biofertilizer#cite_note-4http://en.wikipedia.org/wiki/Biofertilizer#cite_note-4http://en.wikipedia.org/wiki/Fehttp://en.wikipedia.org/wiki/Aluminiumhttp://en.wikipedia.org/wiki/Calciumhttp://en.wikipedia.org/wiki/Organic_acidshttp://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/DNAhttp://en.wikipedia.org/wiki/Molecular_cloninghttp://en.wikipedia.org/wiki/DNA_sequencehttp://en.wikipedia.org/wiki/Nucleotideshttp://en.wikipedia.org/wiki/DNA_replicationhttp://en.wikipedia.org/wiki/DNA_replicationhttp://en.wikipedia.org/wiki/DNA_replicationhttp://en.wikipedia.org/wiki/DNA_replicationhttp://en.wikipedia.org/wiki/Nucleotideshttp://en.wikipedia.org/wiki/DNA_sequencehttp://en.wikipedia.org/wiki/Molecular_cloninghttp://en.wikipedia.org/wiki/DNAhttp://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/Organic_acidshttp://en.wikipedia.org/wiki/Calciumhttp://en.wikipedia.org/wiki/Aluminiumhttp://en.wikipedia.org/wiki/Fehttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-4http://en.wikipedia.org/wiki/Inorganic_phosphatehttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-3http://en.wikipedia.org/wiki/Pseudomonas_putidahttp://en.wikipedia.org/wiki/Pantoea_agglomeranshttp://en.wikipedia.org/wiki/Phosphate_solubilizing_bacteriahttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-2http://en.wikipedia.org/wiki/Azollahttp://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Aulosirahttp://en.wikipedia.org/w/index.php?title=Tolypothrix&action=edit&redlink=1http://en.wikipedia.org/wiki/Anabaenahttp://en.wikipedia.org/wiki/Nostochttp://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Cyanobacteriahttp://en.wikipedia.org/wiki/Blue_green_algaehttp://en.wikipedia.org/wiki/Sugarcanehttp://en.wikipedia.org/wiki/Maizehttp://en.wikipedia.org/wiki/Milletshttp://en.wikipedia.org/wiki/Sorghumhttp://en.wikipedia.org/w/index.php?title=Azospirillum&action=edit&redlink=1http://en.wikipedia.org/wiki/Cottonhttp://en.wikipedia.org/wiki/Mustard_planthttp://en.wikipedia.org/wiki/Maizehttp://en.wikipedia.org/wiki/Wheathttp://en.wikipedia.org/wiki/Azotobacterhttp://en.wikipedia.org/wiki/Rhizobiumhttp://en.wikipedia.org/wiki/Blue_green_algaehttp://en.wikipedia.org/wiki/Azotobacterhttp://en.wikipedia.org/wiki/Rhizobiumhttp://en.wikipedia.org/wiki/Chemical_fertilizershttp://en.wikipedia.org/wiki/Soil_fertilityhttp://en.wikipedia.org/wiki/Plant-growth_promoting_rhizobacteriahttp://en.wikipedia.org/wiki/Pesticideshttp://en.wikipedia.org/wiki/Chemical_fertilizershttp://en.wikipedia.org/wiki/Phosphorushttp://en.wikipedia.org/wiki/Nitrogen_fixationhttp://en.wikipedia.org/wiki/Biofertilizer#cite_note-vessey2003-1http://en.wikipedia.org/wiki/Rhizospherehttp://en.wikipedia.org/wiki/Microorganism7/29/2019 DNA Technology and Forensics
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Contents
1 Introduction
2 Creating recombinant DNA
3 Expression of recombinant DNA 4 Properties of organisms containing recombinant DNA
5 Applications of recombinant DNA technology
6 History of recombinant DNA
7 Controversy
8 See also
9 References
o 9.1 Further reading
10 External links
Introduction
Recombinant DNA molecules are sometimes called chimeric DNA, because they are usually
made of material from two different species, like the mythical chimera. R-DNA technology uses
palindromic sequences and leads to the production ofsticky and blunt ends.
The DNA sequences used in the construction of recombinant DNA molecules can originate from
any species. For example, plant DNA may be joined to bacterial DNA, or human DNA may bejoined with fungal DNA. In addition, DNA sequences that do not occur anywhere in nature may
be created by the chemical synthesis of DNA, and incorporated into recombinant molecules.
Using recombinant DNA technology and synthetic DNA, literally any DNA sequence may be
created and introduced into any of a very wide range of living organisms.
Proteins that result from the expression of recombinant DNA within living cells are termed
recombinant proteins. When recombinant DNA encoding a protein is introduced into a host
organism, the recombinant protein will not necessarily be produced.[citation needed]
Expression of
foreign proteins requires the use of specialized expression vectors and often necessitates
significant restructuring of the foreign coding sequence.[citation needed]
Recombinant DNA differs from genetic recombination in that the former results from artificialmethods in the test tube, while the latter is a normal biological process that results in the
remixing of existing DNA sequences in essentially all organisms.
Creating recombinant DNA
Main article: Molecular cloning
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Construction of recombinant DNA, in which a foreign DNA fragment is inserted into a plasmid vector. In
this example, the gene indicated by the white color is inactivated upon insertion of the foreign DNAfragment.
Molecular cloning is the laboratory process used to create recombinant DNA.[1][2][3][4]
It is one of
two widely-used methods (along with polymerase chain reaction, abbr. PCR) used to direct the
replication of any specific DNA sequence chosen by the experimentalist. The fundamentaldifference between the two methods is that molecular cloning involves replication of the DNA
within a living cell, while PCR replicates DNA in the test tube, free of living cells.
Formation of recombinant DNA requires a cloning vector, a DNA molecule that will replicate
within a living cell. Vectors are generally derived from plasmids orviruses, and represent
relatively small segments of DNA that contain necessary genetic signals for replication, as wellas additional elements for convenience in inserting foreign DNA, identifying cells that contain
recombinant DNA, and, where appropriate, expressing the foreign DNA. The choice of vector
for molecular cloning depends on the choice of host organism, the size of the DNA to be cloned,
and whether and how the foreign DNA is to be expressed.[5]
The DNA segments can becombined by using a variety of methods, such as restriction enzyme/ligase cloning orGibson
assembly.
In standard cloning protocols, the cloning of any DNA fragment essentially involves seven steps:
(1) Choice of host organism and cloning vector, (2) Preparation of vector DNA, (3) Preparation
of DNA to be cloned, (4) Creation of recombinant DNA, (5) Introduction of recombinant DNA
into the host organism, (6) Selection of organisms containing recombinant DNA, (7) Screeningfor clones with desired DNA inserts and biological properties.
[4]These steps are described in
some detail in a related article (molecular cloning).
Expression of recombinant DNA
Main article: Gene expression
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Following transplantation into the host organism, the foreign DNA contained within therecombinant DNA construct may or may not be expressed. That is, the DNA may simply be
replicated without expression, or it may be transcribed and translated so that a recombinantprotein is produced. Generally speaking, expression of a foreign gene requires restructuring the
gene to include sequences that are required for producing a mRNA molecule that can be used by
the host's translational apparatus (e.g. promoter, translational initiation signal, and transcriptionalterminator).[6]
Specific changes to the host organism may be made to improve expression of theectopic gene. In addition, changes may be needed to the coding sequences as well, to optimize
translation, make the protein soluble, direct the recombinant protein to the proper cellular or
extracellular location, and stabilize the protein from degradation.[7]
Properties of organisms containing recombinant DNA
In most cases, organisms containing recombinant DNA have apparently normal phenotypes. Thatis, their appearance, behavior and metabolism are usually unchanged, and the only way to
demonstrate the presence of recombinant sequences is to examine the DNA itself, typically using
a polymerase chain reaction (PCR) test.[8]
Significant exceptions exist, and are discussed below.
If the rDNA sequences encode a gene that is expressed, then the presence of RNA and/or protein
products of the recombinant gene can be detected, typically using RT-PCRorwesternhybridization methods.
[8]Gross phenotypic changes are not the norm, unless the recombinant
gene has been chosen and modified so as to generate biological activity in the host organism.[9]
Additional phenotypes that are encountered include toxicity to the host organism induced by therecombinant gene product, especially if it is over-expressed or expressed within inappropriate
cells or tissues.
In some cases, recombinant DNA can have deleterious effects even if it is not expressed. One
mechanism by which this happens is insertional inactivation, in which the rDNA becomesinserted into a host cells gene. In some cases, researchers use this phenomenon to knock out
genes in order to determine their biological function and importance.[10]
Another mechanism bywhich rDNA insertion into chromosomal DNA can affect gene expression is by inappropriate
activation of previously unexpressed host cell genes. This can happen, for example, when a
recombinant DNA fragment containing an active promoter becomes located next to a previouslysilent host cell gene, or when a host cell gene that functions to restrain gene expression
undergoes insertional inactivation by recombinant DNA.
Applications of recombinant DNA technology
Recombinant DNA is widely used in biotechnology, medicine and research. Today, recombinantproteins and other products that result from the use of rDNA technology are found in essentiallyevery western pharmacy, doctor's or veterinarian's office, medical testing laboratory, and
biological research laboratory. In addition, organisms that have been manipulated using
recombinant DNA technology, and products derived from those organisms have found their way
into many farms, supermarkets, home medicine cabinets and even pet shops.
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The most common application of recombinant DNA is in basic research, where it is important to
most current work in the biological and biomedical sciences.[8]
Recombinant DNA is used to
identify, map and sequence genes, and to determine their function. rDNA probes are employed inanalyzing gene expression within individual cells, and throughout the tissues of whole
organisms. Recombinant proteins are widely used as reagents in laboratory experiments and to
generate antibody probes for examining protein synthesis within cells and organisms.
[2]
Many additional practical applications of recombinant DNA are found in industry, food
production, human and veterinary medicine, in agriculture, and in bioengineering.[2]
Somespecific examples are identified below.
Recombinant chymosin
found in rennet, is an enzyme required to manufacture cheese. It was the first genetically
engineered food additive to be used commercially. Traditionally, processors obtained chymosin
from rennet, a preparation derived from the fourth stomach of milk-fed calves. Scientists
engineered a non-pathogenic strain (K-12) ofE. colibacteria for large-scale laboratoryproduction of the enzyme. This microbiologically produced recombinant enzyme, identical
structurally to the calf derived enzyme, costs less and is produced in abundant quantities. Today
about 60% of U.S. hard cheese is made with genetically engineered chymosin. In 1990, FDA
granted chymosin "generally-recognized-as-safe" (GRAS) status based on data showing that the
enzyme was safe.[11]
Recombinant human insulin
almost completely replaced insulin obtained from animal sources (e.g. pigs and cattle) for the
treatment of insulin-dependent diabetes. A variety of different recombinant insulin preparations
are in widespread use.[12]Recombinant insulin is synthesized by inserting the human insulin
gene intoE. coli, which then produces insulin for human use.[13]
Recombinant human growth hormone (HGH, somatotropin)
administered to patients whose pituitary glands generate insufficient quantities to support
normal growth and development. Before recombinant HGH became available, HGH for
therapeutic use was obtained from pituitary glands of cadavers. This unsafe practice led to some
patients developing Creutzfeldt-Jacob disease. Recombinant HGH eliminated this problem, and
is now used therapeutically.[14]
It has also been misused as a performance enhancing drug by
athletes and others.[15]
DrugBank entry
Recombinant blood clotting factor VIII
a blood-clotting protein that is administered to patients with forms of the bleeding disorder
hemophilia, who are unable to produce factor VIII in quantities sufficient to support normal
blood coagulation.[16]Before the development of recombinant factor VIII, the protein was
obtained by processing large quantities of human blood from multiple donors, which carried a
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very high risk of transmission ofblood borne infectious diseases, for example HIV and hepatitis
B. DrugBank entry
Recombinant hepatitis B vaccine
prevention ofhepatitis B infection is controlled through the use of a recombinant hepatitis B
vaccine, which contains a form of the hepatitis B virus surface antigen that is produced in yeast
cells. The development of the recombinant subunit vaccine was an important and necessary
development because hepatitis B virus, unlike other common viruses such as polio virus, cannot
be grown in vitro. Vaccine information from Hepatitis B Foundation
Diagnosis of infection with HIV
each of the three widely-used methods for diagnosing HIV infection has been developed using
recombinant DNA. The antibody test (ELISA or western blot) uses a recombinant HIV protein to
test for the presence ofantibodies that the body has produced in response to an HIV infection.
The DNA test looks for the presence of HIV genetic material using reverse transcriptasepolymerase chain reaction (RT-PCR). Development of the RT-PCR test was made possible by the
molecular cloning and sequence analysis of HIV genomes. HIV testing page from US Centers for
Disease Control (CDC)
Golden rice
a recombinant variety of rice that has been engineered to express the enzymes responsible for
-carotene biosynthesis.[9]This variety of rice holds substantial promise for reducing the
incidence ofvitamin A deficiency in the world's population.[17]
Golden rice is not currently in use,
pending the resolution of intellectual property, environmental and nutritional issues.
Herbicide-resistant crops
commercial varieties of important agricultural crops (including soy, maize/corn, sorghum,
canola, alfalfa and cotton) have been developed which incorporate a recombinant gene that
results in resistance to the herbicide glyphosate (trade name Roundup), and simplifies weed
control by glyphosate application.[18]
These crops are in common commercial use in several
countries.
Insect-resistant crops
Bacillus thuringeiensis is a bacterium that naturally produces a protein (Bt toxin) withinsecticidal properties.[17]The bacterium has been applied to crops as an insect-control strategy
for many years, and this practice has been widely adopted in agriculture and gardening.
Recently, plants have been developed which express a recombinant form of the bacterial
protein, which may effectively control some insect predators. Environmental issues associated
with the use of these transgenic crops have not been fully resolved.[19]
http://en.wikipedia.org/wiki/Blood-borne_diseasehttp://www.drugbank.ca/molecules/19http://en.wikipedia.org/wiki/Hepatitis_B_vaccinehttp://en.wikipedia.org/wiki/Hepatitis_Bhttp://en.wikipedia.org/wiki/Polio_virushttp://en.wikipedia.org/wiki/In_vitrohttp://www.hepb.org/hepb/vaccine_information.htmhttp://en.wikipedia.org/wiki/HIVhttp://en.wikipedia.org/wiki/HIV_testhttp://en.wikipedia.org/wiki/ELISAhttp://en.wikipedia.org/wiki/Western_blothttp://en.wikipedia.org/wiki/Antibodieshttp://en.wikipedia.org/wiki/RT-PCRhttp://www.cdc.gov/hiv/topics/testing/index.htmhttp://www.cdc.gov/hiv/topics/testing/index.htmhttp://en.wikipedia.org/wiki/Golden_ricehttp://en.wikipedia.org/wiki/%CE%92-carotenehttp://en.wikipedia.org/wiki/%CE%92-carotenehttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C10634784-9http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C10634784-9http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C10634784-9http://en.wikipedia.org/wiki/Vitamin_A_deficiencyhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Glyphosatehttp://en.wikipedia.org/wiki/Glyphosatehttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C16916934-18http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C16916934-18http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C16916934-18http://en.wikipedia.org/wiki/Bacillus_thuringiensishttp://en.wikipedia.org/wiki/Bt_toxinhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Transgenichttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C12949561-19http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C12949561-19http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C12949561-19http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C12949561-19http://en.wikipedia.org/wiki/Transgenichttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Bt_toxinhttp://en.wikipedia.org/wiki/Bacillus_thuringiensishttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C16916934-18http://en.wikipedia.org/wiki/Glyphosatehttp://en.wikipedia.org/wiki/Glyphosatehttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C15793573-17http://en.wikipedia.org/wiki/Vitamin_A_deficiencyhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C10634784-9http://en.wikipedia.org/wiki/%CE%92-carotenehttp://en.wikipedia.org/wiki/Golden_ricehttp://www.cdc.gov/hiv/topics/testing/index.htmhttp://www.cdc.gov/hiv/topics/testing/index.htmhttp://en.wikipedia.org/wiki/RT-PCRhttp://en.wikipedia.org/wiki/Antibodieshttp://en.wikipedia.org/wiki/Western_blothttp://en.wikipedia.org/wiki/ELISAhttp://en.wikipedia.org/wiki/HIV_testhttp://en.wikipedia.org/wiki/HIVhttp://www.hepb.org/hepb/vaccine_information.htmhttp://en.wikipedia.org/wiki/In_vitrohttp://en.wikipedia.org/wiki/Polio_virushttp://en.wikipedia.org/wiki/Hepatitis_Bhttp://en.wikipedia.org/wiki/Hepatitis_B_vaccinehttp://www.drugbank.ca/molecules/19http://en.wikipedia.org/wiki/Blood-borne_disease7/29/2019 DNA Technology and Forensics
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History of recombinant DNA
Main article: History of biotechnology
The idea for recombinant DNA was first proposed by Peter Lobban, a graduate student of Prof.Dale Kaiser in the Biochemistry Department at Stanford University Medical School.
[20]The first
publications describing the successful production and intracellular replication of recombinantDNA appeared in 1972 and 1973.
[21][22][23]Stanford University applied for a US patent on
recombinant DNA in 1974, listing the inventors as Stanley N. Cohen and Herbert W. Boyer; this
patent was awarded in 1980.[24]
The first licensed drug generated using recombinant DNAtechnology was human insulin, developed by Genentech and Licensed by Eli Lilly and
Company.[25]
Controversy
Scientists associated with the initial development of recombinant DNA methods recognized that
the potential existed for organisms containing recombinant DNA to have undesirable or
dangerous properties. At the 1975 Asilomar Conference on Recombinant DNA, these concernswere discussed and a voluntary moratorium on recombinant DNA research was initiated for
experiments that were thought to be particularly risky. This moratorium was widely observeduntil the National Institutes of Health (USA) developed and issued formal guidelines for rDNA
work. Today, recombinant DNA molecules and recombinant proteins are usually not regarded as
dangerous. However, concerns remain about some organisms that express recombinant DNA,
particularly when they leave the laboratory and are introduced into the environment or foodchain. These concerns are discussed in the articles on genetically-modified organisms and
genetically-modified food controversies.
http://en.wikipedia.org/wiki/History_of_biotechnologyhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-20http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-20http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-20http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4342968-21http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4342968-21http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4594039-23http://en.wikipedia.org/wiki/Stanford_Universityhttp://en.wikipedia.org/wiki/Stanley_N._Cohenhttp://en.wikipedia.org/wiki/Herbert_W._Boyerhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C11810894-24http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C11810894-24http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C11810894-24http://en.wikipedia.org/wiki/Genentechhttp://en.wikipedia.org/wiki/Eli_Lilly_and_Companyhttp://en.wikipedia.org/wiki/Eli_Lilly_and_Companyhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C6337396-25http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C6337396-25http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C6337396-25http://en.wikipedia.org/wiki/Asilomar_Conference_on_Recombinant_DNAhttp://en.wikipedia.org/wiki/Genetically_modified_organismhttp://en.wikipedia.org/wiki/Genetically_modified_food_controversieshttp://en.wikipedia.org/wiki/Genetically_modified_food_controversieshttp://en.wikipedia.org/wiki/Genetically_modified_organismhttp://en.wikipedia.org/wiki/Asilomar_Conference_on_Recombinant_DNAhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C6337396-25http://en.wikipedia.org/wiki/Eli_Lilly_and_Companyhttp://en.wikipedia.org/wiki/Eli_Lilly_and_Companyhttp://en.wikipedia.org/wiki/Genentechhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C11810894-24http://en.wikipedia.org/wiki/Herbert_W._Boyerhttp://en.wikipedia.org/wiki/Stanley_N._Cohenhttp://en.wikipedia.org/wiki/Stanford_Universityhttp://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4594039-23http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4342968-21http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-pmid.7C4342968-21http://en.wikipedia.org/wiki/Recombinant_DNA#cite_note-20http://en.wikipedia.org/wiki/History_of_biotechnology