文献类型：期刊文献

中文题名：基因atpA、purHD和ndh的过量表达对大肠杆菌DH5α生长的影响

英文题名：Effects of overexpression of atpA,purHD and ndh on the growth of Escherichia coli DH5α

作者：张艳军[1];张晓云[1];李志敏[1];叶勤[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2008

卷号：48

期号：8

起止页码：1042

中文期刊名：微生物学报

外文期刊名：Acta Microbiologica Sinica

收录：CSTPCD;;Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；PubMed;

基金：国家高技术研究发展计划项目(2002AA217021);上海市重点学科建设项目(B505)~~

语种：中文

中文关键词：大肠杆菌;表达;atpA;purH;ndh

外文关键词：Escherichia coli; Expression; atpA; purH; ndh

摘要：【目的】理解大肠杆菌DH5α及其耐乙酸突变株DA19在基本培养基中生长和乙酸积累差异的机理。【方法】根据前期针对两个菌株蛋白质组和培养中物料及能量平衡的分析,通过PCR分别扩增大肠杆菌DH5α及DA19的atpIBEFHAGDC基因、purHD基因和与NADH氧化代谢相关ndh基因及其各自的调节区,以及NADH脱氢酶Ⅰ基因(nuoA-N)的调节区。将atpA、purHD和ndh基因分别克隆到载体pTrc99a中,转化DH5α,研究基因过量表达对生长的影响。【结果】测序结果表明两个菌株的这些DNA序列与公布的大肠杆菌K-12的相应序列完全一致,在DH5α菌株中分别过量表达atpA、purHD或ndh基因可以在一定程度上改善菌体的生长。【结论】过量表达purHD或ndh基因比过量表达atpA基因的效果更好,但与DA19的生长相比仍然存在较大差距,反映了代谢全局调节的重要性。
[Objective] To study the mechanism of the differences in cell growth and acetate production between Escherichia coli DH5α and its acetate-tolerant mutant DA19. [Methods] Based on our previous analysis of the proteome and mass and energy balances for both strains, we first amplified the genes of atpIBEFHAGDC, putriD, and ndh, their regulation regions, as well as the regulation region of nuoA-N in both strains by PCR, and compared the sequences. Then, we constructed the recombinant strains DH5α （pTrc99a-atpA）, DH5α（pTrc99a-purHD） and DH5α （pTrc99a-ndh） to study the effect of overexpression of atpA, purHD and ndh in DH5α on growth. [Results] The sequences of the structural genes and the regulation regions in the two strains were completely identical with those of E. coli K-12. Overexpression of atpA, purHD and ndh separately in DH5α improved cell growth to some extent. [Conclusion] Overexpression of ndh, purHD and atpA resulted in improved growth of DH5α, but the growth of these recombinant strains was still lower than that of DA19, indicating the importance of global regulation to the cellular metabolism.