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Microbial Genetics (Micr340) . Lecture 13 Molecular Basis of Recombination. Two Types of Recombination. Nonhomologous recombination Occurs very rarely Requires special proteins Homologous recombination Occurs much more often Occurs between same or very similar sequences.
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Microbial Genetics (Micr340) Lecture 13 Molecular Basis of Recombination
Two Types of Recombination • Nonhomologous recombination • Occurs very rarely • Requires special proteins • Homologous recombination • Occurs much more often • Occurs between same or very similar sequences
Recombination requirements • Identical or very similar sequences in the cross-over region • Complementary base pairing between double-stranded DNA molecules • Recombination enzymes • Heteroduplex formation
Models of recombination • Holliday double-strand invasion model
Single-strand Invasion Model
Molecular Basis for Recombination in E. coli • chi () Sites and the RecBCD nuclease • Synapse Formation and the RecA Protein • The Ruv and RecG proteins and the migration and cutting of Holliday Junctions
Phage Recombination Pathways • Rec Proteins and Phages T4 and T7 • Gene 49 protein of T4 and gene 3 protein of T7 are similar to RuvC • Gene 46 and 47 products are similar to RecBCD • The RecE pathway of the rac prophage • RecE protein can substitute for RecBCD nuclease • The phage lred system • Red system requires l genes exo and bet. • Does not require RecA protein
Genetic Analysis of Recombination in Bacteria • Isolating Rec- mutants in E. coli • Other recombination genes • Gene conversion and other manifestations of heteroduplex formation during recombination