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lec15----- - Announcements Quiz 4 due today Regrades due...

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Announcements ± Quiz 4: due today ± Regrades: due Wed. Recombinant DNA technology and Gene cloning
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Early studies of lambda led the way to recombinant DNA technology Cos sites are 12 bases long pGGGCGGCGACCTC G G CCCCGCCGCTGGAp 100,000 BP 5’ 3’ CCCCGCCGCTGGAG GGGGCGGCGACCTC
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Could cohesive ends allow one to bring two different DNAs together? ± How could cohesive ends be made? ± Restriction/modification system was discovered again from bacteriophage studies. Observation: Phage grown on a particular strain of E. coli , could infect same strain and give high yields of progeny High yield (100’s phage)
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Phage grown in different strain of E. coli would give low yield of progeny Low yield (no phage) Restriction/Modification Phage DNA was cleaved into fragments
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Host DNA was protected from cleavage by modifying DNA with methylation Not cleaved by Restriction enzymes If phage DNA was methylated at correct sequences, the DNA was protected from restriction cutting, yielding high numbers of progeny High yield (100’s phage)
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correct sequences, the DNA was cut with restriction enzyme and very few progeny were formed Low yield (no phage) Restriction/modification system ± Primitive immune system ± Protected bacteria from viruses ± Each strain of bacteria had different
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