XU Xu-dong, KONG Ren-qiu, LIU Ke, WANG Jiang-xin, NING De-gang. PCR AMPLIFICATIONS OF FLANKING SEQUENCES OF ORFs IN THE GENOME OF SYNECHOCYSTIS SP. PCC6803 AND A STRATEGY FOR TARGETED GENE DISRUPTION[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(6): 544-550.
Citation: XU Xu-dong, KONG Ren-qiu, LIU Ke, WANG Jiang-xin, NING De-gang. PCR AMPLIFICATIONS OF FLANKING SEQUENCES OF ORFs IN THE GENOME OF SYNECHOCYSTIS SP. PCC6803 AND A STRATEGY FOR TARGETED GENE DISRUPTION[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(6): 544-550.

PCR AMPLIFICATIONS OF FLANKING SEQUENCES OF ORFs IN THE GENOME OF SYNECHOCYSTIS SP. PCC6803 AND A STRATEGY FOR TARGETED GENE DISRUPTION

  • Received Date: July 09, 2001
  • Rev Recd Date: July 20, 2001
  • Published Date: November 24, 2001
  • The flanking sequences of 1927 potential protein-coding genes of Synechocystis sp. PCC6803 were amplified by polymerase chain reactions. The PCR products for the sll0267-sll0268-sll0269 region in the genomes of 4 substrains were inconsistent with those predicted by Kazusa DNA database. As shown with chlorophyll synthesis genes chlH and chlL as the examples, products of 3-piece-ligation PCR can be effectively applied to targeted gene disruptions in the genome of Synechocystis sp. PCC6803.
  • [1]
    Anderson SL, McIntosh L. Light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. strain PCC6803;a blue-light-requiring process [J]. J Bacteriol 1991, 173:2761-2767[2] Kaneko T, Sato S, Kotani H, et al. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions [J]. DNA Res 1996, 3:109-136[3] Shintani D, DellaPenna D. Elevating the vitamin E content of plants through metabolic engineering [J]. Science, 1998, 282:2098-2100[4] 孔任秋,徐旭东,冯勃,集胞藻的随机插入诱变和光激活异养突变株的筛选[J]. 水生生物学报,2001,25(6):537-543[5] Elhai J, Wolk CP. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers [J]. Gene, 1988, 68:119-138[6] Kamei A, Yuasa T, Orikawa K, et al. A eukaryotic-type protein kinase, SpkA, is required for normal motility of unicellular cyanobacterium Synechocystis sp. strain PCC6803 [J]. J Bacteriol, 2001, 183:1505-1510[7] Okamoto S, Ikeuchi M, Ohmori M. Experimental analysis of recently transposed insertion sequences in the cyanobacterium Synechocystis sp. PCC6803 [J]. DNA Res, 1999, 6:265-273[8] Murphy KC, Campellone KG, Poteete AR. PCR-mediated gene replacement in Escherichia coli. Gene [J], 2000, 246:321-330[9] Taroncher-Oldenburg G, Stephanopoulos G. Targeted, PCR-based gene disruption in cyanobacteria;inactivation of the polyhydroxyalkanoic acid synthase genes in Synechocystis sp. PCC6803 [J]. Appl Microbiol Biotechnol, 2000, 54:677-680[10] Suzuki JY, Bollivar DW, Bauer CE. Genetic analysis of chlorophyll biosynthesis [J]. Ann Rev Genet, 1997, 31:61-89[11] Wu Q, Vermaas WFJ. Light-dependent chlorophyll a biosynthesis upon chlL deletion in wild type and photosystem I-less strains of the cyanobacterium Synechocystis sp. PCC6803 [J]. Plant Mol Biol, 1995, 29:933-945

    Anderson SL, McIntosh L. Light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. strain PCC6803;a blue-light-requiring process [J]. J Bacteriol 1991, 173:2761-2767[2] Kaneko T, Sato S, Kotani H, et al. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions [J]. DNA Res 1996, 3:109-136[3] Shintani D, DellaPenna D. Elevating the vitamin E content of plants through metabolic engineering [J]. Science, 1998, 282:2098-2100[4] 孔任秋,徐旭东,冯勃,集胞藻的随机插入诱变和光激活异养突变株的筛选[J]. 水生生物学报,2001,25(6):537-543[5] Elhai J, Wolk CP. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers [J]. Gene, 1988, 68:119-138[6] Kamei A, Yuasa T, Orikawa K, et al. A eukaryotic-type protein kinase, SpkA, is required for normal motility of unicellular cyanobacterium Synechocystis sp. strain PCC6803 [J]. J Bacteriol, 2001, 183:1505-1510[7] Okamoto S, Ikeuchi M, Ohmori M. Experimental analysis of recently transposed insertion sequences in the cyanobacterium Synechocystis sp. PCC6803 [J]. DNA Res, 1999, 6:265-273[8] Murphy KC, Campellone KG, Poteete AR. PCR-mediated gene replacement in Escherichia coli. Gene [J], 2000, 246:321-330[9] Taroncher-Oldenburg G, Stephanopoulos G. Targeted, PCR-based gene disruption in cyanobacteria;inactivation of the polyhydroxyalkanoic acid synthase genes in Synechocystis sp. PCC6803 [J]. Appl Microbiol Biotechnol, 2000, 54:677-680[10] Suzuki JY, Bollivar DW, Bauer CE. Genetic analysis of chlorophyll biosynthesis [J]. Ann Rev Genet, 1997, 31:61-89[11] Wu Q, Vermaas WFJ. Light-dependent chlorophyll a biosynthesis upon chlL deletion in wild type and photosystem I-less strains of the cyanobacterium Synechocystis sp. PCC6803 [J]. Plant Mol Biol, 1995, 29:933-945

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