运动发酵单胞菌限制-修饰系统缺失突变株的构建及其性质分析(英文)  

Construction and Characterization of Restriction-Modification Deficient Mutants in Zymomonas mobilis ZM4

作　　者：吴波[1] 何明雄[2,3] 冯红[1] 张艳[1] 胡启春[2,3] 张义正[1] 

机构地区：[1]四川大学生命科学学院,四川省分子生物学与生物技术重点实验室,生物资源与生态环境教育部重点实验室,成都610064 [2]农业部沼气科学研究所生物质能技术研究中心,成都610041 [3]农业部农村可再生能源开发与利用重点实验室,成都610041

出　　处：《应用与环境生物学报》2013年第2期189-197,共9页Chinese Journal of Applied and Environmental Biology

基　　金：the National Natural Science Foundation of China(No.31000028);the Sichuan Key Technology R&D Program(No.2009NZ00045);the Sci-Tech Fund Project of Chinese Academy of Agricultural Sciences(2009and 2011);the National Science and Technology Pillar Program of China(No.2007BAC18B04)~~

摘　　要：低遗传转化率阻碍了产乙醇模式菌——运动发酵单胞菌的遗传或分子生物学操作.本研究将运动发酵单胞菌菌株ZM4的5个限制-修饰系统相关基因失活,构建了5个限制-修饰系统突变菌株.研究结果表明:在突变株Zmmrr和Zm1933中,甲基化穿梭表达质粒pBBR1MCS-tet的电转化率分别提高了17倍和2倍,而限制修饰基因ZMO0575的失活则明显降低了甲基化修饰质粒和非甲基化质粒的转化率.较之其它3个突变株,突变株Zmmrr和Zm1933具有更高的遗传稳定性.发酵实验结果进一步表明这些限制-修饰系统突变株并未显著改变运动发酵单胞菌的主要性质,例如细胞生长、葡萄糖利用率和乙醇产量等.研究运动发酵单胞菌的限制-修饰系统将有助于构建适合于分子遗传操作的基因工程菌株.Low transformation efficiency is an obstacle to genetic or molecular manipulations in ethanologen Zymomonas mobilis. In the present study, 5 defective strains were constructed in Z. mobilis strain ZM4 by inactivating restriction-modification （R-M） system candidate genes. Inactivation of ZMO0028 （mrr） and ZMO1933 significantly improved electroporation efficiency by 17 folds and 2 folds when ZM4 was transformed with the methylated plasmid DNA. Disruption of ZMO0575 significantly decreased the transformation efficiency when transformed with both methylated and unmethylated plasmid DNAs. In comparison with other mutants, Zmmrr and Zm1933 displayed high stability. Furthermore, fermentation results showed that R-M mutants did not significantly alter the major bacterial traits such as growth, glucose utilization and ethanol yield. In conclusion, R-M systems in Z. mobilis were investigated in this study, and the characterization of those R-M genes contributed to creating engineering strains suitable for genetic and molecular manipulations. Fig 5, Tab 3, Ref 37

关 键 词：运动发酵单胞菌 限制-修饰系统 基因敲除 电转化 乙醇发酵 

分 类 号：Q93[生物学—微生物学]