DNA recombinant technology & genetic engineering (1) several concepts

1. DNA recombinant technology & genetic engineering (1) several concepts * recombinant DNA * technology of recombinant DNA

2. * Genetic engineering • general procedure • A. separate • B. cut • C. linkage • D. transfection or • transformation

3. E. Screening a. separate * Vectors concept type

10. * the sources of foreign target DNA three methods genomic DNA complementary DNA,cDNA synthetic DNA fragment Lehninger 112601

12. Genomic DNA library cDNA library • b. cut • restriction endonuclease • concept • nomination • type

15. 5’ G-A-A-T-T-C 3’ 3’ C-T-T-A-A-G 5’ 5’ G 3’ 5’ A-A-T-T-C 3’ 3’ C-T-T-A-A-5’ 3’ G 5’ sticky end

16. 5’-A-G-C-T-3’ 3’-T-C-G-A-5’ 5’-A-G-3’ 5’-C-T-3’ 3’-T-C-5’ 3’-G-A-5’ blunt end

17. Linkage • d. transformation or transfection • microinjection • transgenic • E. Screening • drug-resistance gene • X-gal screening

19. Southern blot *digested with enzyme * electrophoresis * transfer * hybridization * autoradiography

23. Lehninger 92201