多变鱼腥藻ATCC 29413基因的一步插入失活  

ONE-STEP GENE INTERRUPTION IN ANABAENA VARIABILIS ATCC 29413

作　　者：雷棋琴 戢水玲 黄任衡 高宏[1] 徐旭东[1] 孔任秋[1] LEI Qi-Qin;JI Shui-Ling;HUANG Ren-Heng;GAO Hong;XU Xu-Dong;KONG Ren-Qiu(Key Laboratory of Algal Biology,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院水生生物研究所中国科学院藻类生物学重点实验室,武汉430072 [2]中国科学院大学,北京100049

出　　处：《水生生物学报》2021年第1期48-53,共6页Acta Hydrobiologica Sinica

基　　金：现代农业产业技术体系专项资金(CARS-50)资助。

摘　　要：多变鱼腥藻ATCC 29413有潜力发展为异养生长产异形胞蓝藻的研究模式种,但由于细胞内的限制-修饰系统等原因,其基因转移效率极低。研究克隆了该藻株的两个甲基化酶(M.AvaⅠ和M.AvrⅡ)基因ava_3181和ava_4359,构建了辅助质粒pHB6088,对运载质粒进行甲基化保护以避免被细胞中的限制酶切割。以ava_1237和ava_4412基因为例,研究证明利用该辅助质粒和接合转移系统可通过双交换对多变鱼腥藻ATCC 29413的基因实现一步插入失活。There is the potential for Anabaena variabilis ATCC29413 to be developed into a model strain for studies of heterocyst-forming cyanobacteria that can grow heterotrophically.However,partially due to the restriction-modification systems in cells,the efficiency of gene transfer into this strain is extremely low.To this end,we cloned the two methylase genes,ava_3181(M.AvaI)and ava_4359(M.AvrII),of A.variabilis ATCC 29413 and constructed the helper plasmid pHB6088 to protect cargo plasmids from cleavage by restriction enzymes.Taking ava_1237 and ava_4412 as the examples,this study showed that conjugation with this helper plasmid can allow genes to be interrupted in one step through homologous recombination in A.variabilis ATCC 29413.

关 键 词：多变鱼腥藻ATCC 29413 接合转移 辅助质粒 甲基化酶基因 基因插入失活 

分 类 号：Q173[生物学—水生生物学]