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Three-Piece-Ligation PCR and Application in Disruption of Chlorophyll Synthesis Genes in Synechocystis sp. PCC 6803

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Abstract

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.

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Authors and Affiliations

  1. Institute of Hydrobiology, the Chinese Academy of Sciences, Wuhan, Hubei 430072, P.R. China, , , , , , CN

    Renqiu Kong & Xudong Xu

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  1. Renqiu Kong
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  2. Xudong Xu
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Received: 20 June 2001 / Accepted: 30 July 2001

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Kong, R., Xu, X. Three-Piece-Ligation PCR and Application in Disruption of Chlorophyll Synthesis Genes in Synechocystis sp. PCC 6803. Curr Microbiol 44, 241–245 (2002). https://doi.org/10.1007/s00284-001-0032-6

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  • DOI: https://doi.org/10.1007/s00284-001-0032-6

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