lecture_20__05 - Keep in mind that genetic cloning is...

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Keep in mind that genetic cloning is unrelated to the “cloning” of organisms often discussed in the media. Cloning in the media today refers to nuclear transfer. successful with sheep, cows, pigs, rabbits, mice, cats the possibility of doing nuclear transfer with human cells is ethically and politically controversial success rate very low because of : imprinting, chromatin structure, lack of natural signalling events that go on after normal fertilization, etc. Later complications due to telomere shortening and perhaps other unknown causes of aging if nuclear transfer was performed with human DNA, this would be equivalent to asexual reproduction
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Recombinant DNA technology: “Cloning” ~ genetic engineering 1973; Boyer and Cohen realized that bacterial plasmids could be manipulated to include DNA from other plasmids or even other organisms. Once in a plasmid, the DNA would be replicated along with the bacterial chromosome. Therefore, an unlimited supply of the DNA could be made just be growing more and more of the bacterial strain containing the plasmid. Why do we use plasmids to do genetic engineering? plasmids are small (~1,500 to 15,000 bp) they are nonessential
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There are several biochemical tools necessary for cloning: 1. (scissors) Need a mechanism to cut DNA in defined and reproducible ways. Restriction enzymes “Restriction” is a system used by bacteria to remove foreign DNA that has entered the cell Bacterial DNA is methylated by a cellular methylase, while most foreign DNA and bacteriophage DNA is not. Specific endonucleases called restriction enzymes cut the unmethylated but do not cut the methylated DNA. Stryer, Biochemistry, 34-13
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lecture_20__05 - Keep in mind that genetic cloning is...

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