蓝藻的一种人工转化系统研究  

AN ARTIFICIAL TRANSFORMATION SYSTEM OF CYANOBACTERIA

作　　者：闵红涛 王业勤[1] 

机构地区：[1]中国科学院水生生物研究所

出　　处：《水生生物学报》1995年第3期227-233,共7页Acta Hydrobiologica Sinica

摘　　要：本研究用溶菌酶处理ＳｙｎｅｃｈｏｃｏｃｃｕｓＰＣＣ６３０１细胞诱导人工感受态，并用外源质粒ｐＢＲ３２５转化受体细胞表达氯霉素抗性，转化频率接近５×１０￣（－５）转化子／细胞。用转化子ＤＮＡ再进行次级转化，转化频率可达５×１０￣（－４）转化子／细胞。这比以前的研究者对同种藻株，用克隆的ＤＮＡ、通过生理感受态进行转化得到的转化频率要高。ＤＮＡ电泳、次级转化和斑点杂交证明外源质粒通过单交换已经整合到了受体细胞染色体上。这些结果表明，人工转化系统是有效的，并具有可重复性。对于影响转化的一些条件，如ＤＮＡ与细胞保温时间、藻龄、光照或黑暗培养，也同时进行了研究。Artificial competence of Synechococcus PCC 6301 cells was induced by lysozymetreatment and the cells were transformed to chloramphenicol resistance with foreignplasmid pBR325 at a frequency of approximately 5 ×10￣(-5) or 5 × 10￣(-4) with the tra-nsformant DNA. The transformation frequencies were higher than those reported byother workers for the same strain with cloned DNA employing a physiological tran-sformation system.Analyses of DNA electrophoresis,secondary transformation anddot blotting demonstrated that foreign plasmid had integrated into the recipientchromosome by a single crossover event.The results showed that the artificial tran-sformation system was efficient and reproducible. Conditions that affected transfor-mation,such as,incubation time of cells with DNA,age of the cells,light or darkincubation were also studied.

关 键 词：蓝藻 转化 人工转化系统 

分 类 号：Q949.220.3[生物学—植物学]