DNA recombination - Grenetfr：DNA重组- grenetfr.pptVIP

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * DNA replication segregation of homologous chromosomes gametes Chromosome condensation centromer sister chromatids Pairing of homologous chromatids and crossing-over synaptolemal complex 1st mitosis 2ndmitosis Recombination during meiosis * ??Crossing over?? Mitochondrial DNA transmission Exclusive transmission of mother mitochondria simple double homologous sequence frequency : 1/107 base pairs, at least one per chromosome paternal chromosome maternal chromosome Epigenetics Some genes are inactivated by methylation, the methylation state can be transmitted to daughter cells. Example : inactivation of one chromosome X in women Non-Mendelian transmission * ampicillineR blasticidineR target gene blasticidineR Recombination (double crossing-over) WT PHG1A Dphg1a Dphg1b D?phg1a/b PHG1B Anti-PHG1B Anti-PHG1A Benzhegal et al. 2002 Application of recombination : gene knock-out by insertion * 2. Site-specific recombination Condition : presence of a specific sequence repeated twice Mechanism : specialized protein complex, no branch migration Specific case : recombination with a circular DNA molecule Simple recombination Double recombination Recombination with circular DNA : local double recombination (no branch migration) The two states of the bacteriophage l Reversible recombination DNA of the bacterio phage l DNA of E. coli attP attB Recombinant DNA Integrase Integration Host Factor ExcisionaseIntegrase Integration Host Factor Example 1 : site-specific recombination of a virus * Integrase mechanism phage l DNA E. Coli DNA attP attB recombinant DNA pairing, double cleavage, double exchange, ligation * Conformation 1 : phage and bacterial DNA separated Conformation 2 : phage and bacterial DNA fused attB attP bacterial DNA phage DNA * Biswas et al. (2005) A structural basis for allosteric control of DNA recombination by λ integrase Nature 435 : 1059-1066 integratio