Biology 1201A Chapter Notes - Chapter 15: Restriction Fragment Length Polymorphism, Genomic Library, Somatic Cell Nuclear Transfer
15 Jul 2015
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15. 1 dna cloning: gene of interest: gene that researcher wants to study or manipulate, basic research: biological functions, applied research: pharmaceutical & clinical, method: uses bacteria & plasmids- small circular dna molecule that replicate separately from bacterial chromosome. Extract dna from cells containing gene of interest. Cut dna into fragments - > inserted into plasmids. Produces collection of recombinant dna molecules- recombinant dna- dna from 2+ different sources joined together. Identify which bacterium contains plasmid carrying gene of interest & isolate for further study. Dna molecules cut w/ same enzyme: pairing leaves nicks in sugar-phosphate backbone, sealed by ligase: enzyme that has same function in dna replication, fig. Lacz+ gene encodes b-galactosidase - > hydrolyzes sugar lactose & synthetic substrates. Restriction sites are located within lacz+ gene but do not alter gene"s function. Colony produces b-galactosidase- converts x-gal - > blue product.
