Conjugation
I. Bacterial Genetics
Lederberg and Tatum
What was known, what they thought, what it was
Donor (male), recipient (female), pilus, transconjugate
II. Mechanism of Plasmid transfer
Conjugative vs Molibizable
Tra genes
Dtr (DNA transfer & replication)
Mpf (Mating-pair formation)
oriT vs oriV, relaxase, primase, peelaway single strand transfered, replication
Regulation of transfer, FinP (antisense RNA to traJ), FinO stabilizes FinP
Mobilizable plasmids have mob genes which are tra-"like" hitch-hikers
III. Chromosomal Transfer (Bacterial Sex)
Insertion elements
Formation of Recombinant chromosomes
Interspecies mating and barriers to overcome between species
F vs F' vs Hfr
IV. Experiments using conjugation
Testing for conjugational recombination in an Hfr strain
Donated material must have selectable marker
Recipient material should have selectable marker
Filters, growth phases, selection, controls
Gene mapping using conjugation
Gradient of transfer
Time of entry
Complementation using F’s
V. Other mechanisms of plasmid transfer
Chemical
Electroporation
VI. Other systems
Gram positive bacteria
Pheromones
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