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Lecture 15

CAS BI 216 Lecture 15: Genetics - Lecture 15 - Chapter 9

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CAS BI 216
John Celenza

Lecture 15 4417 (Chapter 9) Chapter 9 (cont) Kinds of Vectors o Plasmid 10kbp Ecoli o Cosmid 45 kbp o BAC 100300 kbp o YAC 250 2000 kbp Prokaryotes vs. Eukaryotes o Prokaryotes no nucleus transcription pppNp 5 end middle no introns 3 end stemloop translation ribosome binding site UAAGGAGGAU o shivedalgamo sequence o Eukaryotes nucleus transcription nucleus translation cytoplasm o transcription 7meGpppN 5 cap introns poly A tail translation ribosome binding site 5 cap cDNA CloningcDNA Libraries o cDNA = complementary (to mRNA) o steps isolate cell (liver) isolate mRNA on polydT cellulose column polydT primer primer binds poly A tail mRA TTTTT 5AAAA reverse transcriptase + dNTPs cDNA TTTTT 5AAAA 3 digest away in mRNA DNA self primes DNAP + dNTPs (see notes) o cDNA only mRNA sequence (exons) only from cell type liver genomic DNA library contains all sequences from genome (exons,
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