集胞藻ORFs侧翼序列的PCR扩增和定向基因插入失活策略

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

摘要: 针对集胞藻PCC6803的1927个待定编码基因进行了两侧序列的PCR扩增.4个亚株基因组在.sll0267-sll0268-sll0269区域的PCR扩增产物与KazusaDNA数据存在差异.以叶绿素合成基因chlH和chlL为例,显示三片段连接PCR产物可有效用于集胞藻6803基因组定向插入失活.

Abstract: 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.