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Published byBlaise Elliott Modified over 9 years ago
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DNA Cloning
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Cloning a line of genetically identical cells or individuals derived from a single ancestor produces many copies of a piece of DNA uses a little fraction as gene of interest cultivates a large amount for studying functions
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Functions Biological determine the DNA sequence ultimate information regulation Pharmaceutical manufacture medicines Clinical
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How it Works Combine gene of interest and bacterial plasmid Recombinant DNA DNA from two or more different sources that have been joined together to form a single molecule Amplification and identification
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Zoom In!!! Insertion, the most critical step made possible by restriction endonucleases(aka. restriction enzymes) recognize specific DNA sequences called restriction sites cut the DNA at specific locations Restriction fragments
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Restriction Enzyme defend against viral attack by breaking down the DNA molecules of infecting viruses cleave the sugar-phosphate backbones of DNA to produce sticky ends short single-stranded regions form hydrogen bonds with complementary sticky ends on any other DNA molecules cut with the same enzyme kind of like glue when you fix something
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Cloning Vectors DNA molecules into which a DNA fragment can be inserted to form a recombinant DNA molecule for the purpose of cloning plasmid cloning vectors contains 2 unique genes: amp R gene and lacZ+ gene
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The Two Genes ampR gene encodes an enzyme that breaks down the antibiotic ampicillin lacZ+ gene encodes ß - galactosidase (hydrolyzes the sugar lactorse and synthetic substrates)
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