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Recombinant DNA technology was used to produce large-scale quantities of the human hormone insulin in E. coli as early as 1978. Previously, it was only possible to treat diabetes with pig insulin, which caused allergic reactions in many humans because of differences in the insulin molecule. In addition, human growth hormone (HGH) is used to treat growth disorders in children. The HGH gene was cloned from a cDNA (complementary DNA) library and inserted into E. coli cells by cloning it into a bacterial vector.

Transgenic animals

Although several recombinant proteins used in medicine are successfully produced in bacteria, some proteins need a eukaryotic animal host for proper processing. For this reason, genes have been cloned and expressed in animals such as sheep, goats, chickens, and mice. Animals that have been modified to express recombinant DNA are called transgenic animals ( [link] ).

A photo shows 3 mice under ultraviolet light. All three have white fur that looks purple in the UV light. The middle mouse is non-transgenic and is non-fluorescing. The mice on the left and right are transgenic, and their eyes, ears, nose, and tail fluoresce green under the UV light.
It can be seen that two of these mice are transgenic because they have a gene that causes them to fluoresce under a UV light. The non-transgenic mouse does not have the gene that causes fluorescence. (credit: Ingrid Moen et al.)

Several human proteins are expressed in the milk of transgenic sheep and goats. In one commercial example, the FDA has approved a blood anticoagulant protein that is produced in the milk of transgenic goats for use in humans. Mice have been used extensively for expressing and studying the effects of recombinant genes and mutations.

Transgenic plants

Manipulating the DNA of plants (creating genetically modified organisms, or GMOs) has helped to create desirable traits such as disease resistance, herbicide, and pest resistance, better nutritional value, and better shelf life ( [link] ). Plants are the most important source of food for the human population. Farmers developed ways to select for plant varieties with desirable traits long before modern-day biotechnology practices were established.

A photo of cobs of corn with kernels of varying shape and color.
Corn, a major agricultural crop used to create products for a variety of industries, is often modified through plant biotechnology. (credit: Keith Weller, USDA)

Transgenic plants have received DNA from other species. Because they contain unique combinations of genes and are not restricted to the laboratory, transgenic plants and other GMOs are closely monitored by government agencies to ensure that they are fit for human consumption and do not endanger other plant and animal life. Because foreign genes can spread to other species in the environment, particularly in the pollen and seeds of plants, extensive testing is required to ensure ecological stability. Staples like corn, potatoes, and tomatoes were the first crop plants to be genetically engineered.

Transformation of plants using Agrobacterium tumefaciens

In plants, tumors caused by the bacterium Agrobacterium tumefaciens occur by transfer of DNA from the bacterium to the plant. The artificial introduction of DNA into plant cells is more challenging than in animal cells because of the thick plant cell wall. Researchers used the natural transfer of DNA from Agrobacterium to a plant host to introduce DNA fragments of their choice into plant hosts. In nature, the disease-causing A. tumefaciens have a set of plasmids that contain genes that integrate into the infected plant cell’s genome. Researchers manipulate the plasmids to carry the desired DNA fragment and insert it into the plant genome.

The organic insecticide Bacillus thuringiensis

Bacillus thuringiensis (Bt) is a bacterium that produces protein crystals that are toxic to many insect species that feed on plants. Insects that have eaten Bt toxin stop feeding on the plants within a few hours. After the toxin is activated in the intestines of the insects, death occurs within a couple of days. The crystal toxin genes have been cloned from the bacterium and introduced into plants, therefore allowing plants to produce their own crystal Bt toxin that acts against insects. Bt toxin is safe for the environment and non-toxic to mammals (including humans). As a result, it has been approved for use by organic farmers as a natural insecticide. There is some concern, however, that insects may evolve resistance to the Bt toxin in the same way that bacteria evolve resistance to antibiotics.

Flavrsavr tomato

The first GM crop to be introduced into the market was the FlavrSavr Tomato produced in 1994. Molecular genetic technology was used to slow down the process of softening and rotting caused by fungal infections, which led to increased shelf life of the GM tomatoes. Additional genetic modification improved the flavor of this tomato. The FlavrSavr tomato did not successfully stay in the market because of problems maintaining and shipping the crop.

Section summary

Genetic testing is performed to identify disease-causing genes, and can be used to benefit affected individuals and their relatives who have not developed disease symptoms yet. Gene therapy—by which functioning genes are incorporated into the genomes of individuals with a non-functioning mutant gene—has the potential to cure heritable diseases. Transgenic organisms possess DNA from a different species, usually generated by molecular cloning techniques. Vaccines, antibiotics, and hormones are examples of products obtained by recombinant DNA technology. Transgenic animals have been created for experimental purposes and some are used to produce some human proteins.

Genes are inserted into plants, using plasmids in the bacterium Agrobacterium tumefaciens , which infects plants. Transgenic plants have been created to improve the characteristics of crop plants—for example, by giving them insect resistance by inserting a gene for a bacterial toxin.

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Source:  OpenStax, Concepts of biology. OpenStax CNX. Feb 29, 2016 Download for free at http://cnx.org/content/col11487/1.9
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