engleski

Recombination lullaby: Take #1 (Reachable ends-out recombination) & Take #2 (Elusive multiple integration)

Take 1. Targeted gene replacement via ends-out recombination is an elegant technique by which yeast genome could be easily manipulated using its own recombination potential. DNA fragment containing selectable marker flanked by sequences homologous to the corresponding sidewise regions of the ORF to be deleted undergoes recombination within the cell. Its free, highly recombinogenic ends recombine with the target sequence in the genome resulting in replacement of the endogenous allele with the exogenous one. Take 2. Non-replicative plasmids bearing yeast homology transform the cell only through homologous recombination with the chromosome. This can produce transformants that either have integrated the plasmid molecule into the genome creating at least one functional copy of the targeted locus (gene conversion followed by reciprocal exchange) or replaced the non-functional chromosomal allele with the functional one from the plasmid (gene conversion without reciprocal exchange). However, this process is known to integrate more than one plasmid molecule into the genome leading to multiple plasmid integration.

recombination; ends-out; multiple integration

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