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[1]吴波,何明雄,冯红,等.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][J].应用与环境生物学报,2013,19(02):189-197.[doi:10.3724/SP.J.1145.2013.00189]
 WU Bo,HE Mingxiong,FENG Hong,et al.Construction and Characterization of Restriction-Modification Deficient Mutants in Zymomonas mobilis ZM4[J].Chinese Journal of Applied & Environmental Biology,2013,19(02):189-197.[doi:10.3724/SP.J.1145.2013.00189]
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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]()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
19卷
期数:
2013年02期
页码:
189-197
栏目:
研究论文
出版日期:
2013-04-25

文章信息/Info

Title:
Construction and Characterization of Restriction-Modification Deficient Mutants in Zymomonas mobilis ZM4
作者:
吴波 何明雄 冯红 张艳 胡启春 张义正
"(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)"
Author(s):
WU Bo HE Mingxiong FENG Hong ZHANG Yan HU Qichun ZHANG Yizheng
(1Sichuan Key Laboratory of Molecular Biology & Biotechnology and Key Laboratory of Resource Biology & Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China)
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
Keywords:
"Zymomonas mobilis restriction-modification systems gene knockout electroporation ethanol fermentation"
分类号:
Q933 : TK63
DOI:
10.3724/SP.J.1145.2013.00189
文献标志码:
A
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. 图5 表3 参37
Abstract:
"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"

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备注/Memo

备注/Memo:
国家自然科学基金项目(31000028)、四川省科技支撑计划项目(2009NZ00045)、中国农业科学院科技经费项目(2009和2011)和国家科技支撑计划项目(2007BAC18B04)资助
更新日期/Last Update: 2013-05-02