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BIOL 208: MIDTERM 2 REVIEW.docx

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Department
Biology
Course
BIOL 309
Professor
David Spafford
Semester
Fall

Description
MIDTERM 2 REVIEW Module 6: Cloning and Vectors Transformation: uptake of naked DNA Competency: the ability to take up DNA by transformation E. coli can be induced to take up DNA by: - CaCl2treatment followed by heat shock - Electroporation Ways in which bacteria uptake foreign DNA: a) Conjugation b) Transformation c) Transduction To improve transformation efficiency, use specialized host strains that have mutations in: 1. Endonucleases 2. Recombination genes Bacteriophage: virus that infects bacterial host and exists as a parasite Bacteriophage lambda has linear DNA genome when in capsid Temperate virus Two types of lambda infection: Lysogenic: dormant in host genome Lytic: infects, multiplies, and kills host cell by lysis Types of lambda cloning vectors: a) Insertion vectors: 7.6 kb inserts b) Replacement vectors: 22 kb inserts c) Cosmids: plasmid with cost site -> 32-45 kb of inserts Other types of vectors: a) Expression vectors: recover protein product it encodes b) Transcriptional fusion vectors for bacterial hosts: express coding sequence under control of different promoter c) Translational fusion vectors: insert fused to vector’s promoter or insert fused in-frame to another ORF d) Mammalian expression vectors e) Plant expression vectors f) Supervectors: vectors that would accept large inserts Module 7: Genomic and cDNA Libraries Use genomic libraries to: - Isolate specific pieces of genomic DNA - Isolate fragments adjacent to specific fragment - Gene mapping and genome sequencing Use cDNA libraries to iso
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