24.

3 Biotechnology
Cloning

Cloning is the production of identical copies of an organism This occurs naturally in new plant shoots, bacterial colonies, and identical human twins Gene cloning is the production of identical copies of a gene. Using Recombinant DNA technology Recombinant DNA (rDNA) contains DNA from two or more different sources A vector plasmid (a small accessory ring of DNA in bacteria) or virus is necessary to insert foreign DNA into a host cell Restriction enzymes: cleave the vector DNA and the source DNA at a specific sequence, leaving "sticky" ends, that allow a portion of source DNA to be inserted into the vector DNA

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DNA ligase then seals the openings and recombinant DNA is formed. After recombinant DNA enters a host cell, it may be copied If successful, some particular gene(s) have been cloned and can be recovered Bacterial cells take up recombinant plasmids and clone the new DNA

Reverse Transcription When a viral vector is used, the cloned DNA is inside newly formed bacteriophages To express a human gene in a bacterium, it must not have introns. Reverse transcriptase can be used to make a DNA copy of mRNA; this complementary DNA does not contain introns

Using the Polymerase Chain Reaction

The Polymerase Chain Reaction (PCR) produces many copies of a single gene or piece of DNA. PCR requires DNA polymerase and a supply of nucleotides for the new DNA strands Artificially cycles through a) unzipping DNA, b) copying DNA and c) rezipping DNA Cycle repeats 30-36x so that with 1 starting copy, up to 2^35 new copies may be made.

Mr. Plattener's PCR/ gel electrophoresis (Diagram Package: 24.3 P2 D2~3 Well, its impossible to find these images on the web.)
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DNA Fingerprinting

The entire genome of an individual can be cut by restriction enzymes to yield variable fragment lengths Gel electrophoresis separates fragment lengths Use of probes results in a pattern unique to the individual (sometimes call a DNA fingerprint).

CSI

You can identify a person who has committed a crime, or tell who is related to whom by doing a DNA fingerprint. Since PCR can amplify the smallest amount of DNA, a single sperm, or one cell on a cigarette butt, provide enough DNA to be identified by comparison with sample DNA

Biotechnology

Biotechnology uses natural biological systems to create a product or to achieve an end desired by human being. Transgenic organisms have had a foreign gene inserted into them. (Sorry, the photos of the two transgenic organisms are not provided here to ensure that you feel comfortable reading the notes.)

Who is the criminal?

Transgenic Bacteria

Transgenic Bacteria are used to produce biotechnology products such as: insulin, human growth hormone, t-PA and hepatitis B vaccine. They also add insecticidal toxins to plants, reduce frost damage, degrade wastes, produce chemicals, and help mine metals. Foreign genes are added to protoplasts, plant cells that lack a cell wall, using electric current. Foreign genes in cotton, corn, and potatoes have given the pest resistance; soybeans are made resistant to herbicide for no-till farming

Transgenic Plants cont'd

Transgenic plants produce human hormones, clotting factors, and antibodies in their seeds. One wed has even been engineered to produce plastic granules
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Transgenic Animals

Foreign genes can be inserted into animal eggs by hand or by vortex mixing. Gene pharming uses transgenic farm animals to produce pharmaceuticals in their ilk. There are plans to use animals to produce drugs for the treatment of cystic fibrosis, cancer, blood, diseases, etc. In vitro fertilization is necessary Cloning of mammals was once considered impossible, but has now been accomplished with sheep, calves and goats. A 2ns nucleus from a bioengineered animal is inserted into enucleated (original nucleus removed) eggs from a donor. A surrogate mother gives birth to the cloned animals.

Animal Organs as Biotechnology Products Scientists are genetically engineering pigs to serve as organ donors for human who need transplants. Transplants of organs across species is xenotransplantation. Researchers are trying to make organs less antigenic to humans. One concern is whether pig organs might carry animal viruses into humans; HIV is virus that jumped from monkeys into humans. Tissue engineering is an alternative method of securing transplant material from culturing human tissue from a mixture of cells and synthetic polymers.

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