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Kong, R. et al.

Three-piece-ligation PCR and application in disruption of chlorophyll synthesis genes in Synechocystis sp. PCC 6803.

Linear DNA, consisting of a drug-resistance marker and long flanking sequences, was synthesized by one-step polymerase chain reaction after a three-piece ligating reaction. Chlorophyll synthesis genes, chlH and chlL in Synechocystis sp. PCC 6803, were replaced by a kanamycin-resistance marker through double recombinations with flanking homology regions. Under LAHG conditions, the chlL but not chlH mutant stopped chlorophyll synthesis, while both synthesized chlorophyll in the light.[1]

References

Three-piece-ligation PCR and application in disruption of chlorophyll synthesis genes in Synechocystis sp. PCC 6803. Kong, R., Xu, X. Curr. Microbiol. (2002) [Pubmed]

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