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学科主题: 微生物学
题名: Construction of E. coli NAD+ auxotrophic strains and the biotechnological application thereof
作者: Zhou YJ(周雍进) ;  Hou SH(侯淑华) ;  Zhang SF(张素芳) ;  Zhao ZB(赵宗保)
会议文集: Book of Abstracts
会议名称: Asian Congress on Biotechnology
会议日期: 2011-5-11
出版日期: 2011
会议地点: 上海
通讯作者: 赵宗保
出版者: 待补充
出版地: 待补充
合作性质: 分会口头报告
部门归属: 1816
主办者: 亚洲生物技术联合会
摘要: NAD+ and its reduced form NADH are essential cofactors in biological systems. They function as cofactors in over 300 redox reactions in vivo1. The level of NAD+ and NADH is tightly controlled by a variety of mechanisms including their biosynthesis and salvage. Therefore, it is difficult to answer some fundamental questions such as the minimal NAD+ level for cell growth and the biological consequences of abnormal activity of a specific NAD+-dependent enzyme. We expressed the NTT4 gene from Chlamydiae UWE25 in Escherichia coli BW25113, as the NTT4 protein was reported as a NAD+ transporter that can specifically transport intact NAD+ across cytoplasmic membrane2. We knocked out the nadC gene responsible for de novo biosynthesis of NAD+ and constructed the strain E. coli BW25113 (ΔnadC, NTT4). It was found that NAD+ in the culture media could significantly promote the growth of BW25113 (ΔnadC, NTT4), suggesting that the NTT4 protein was functional in E. coli (Fig A, B). We then disrupted the other two genes, nadD and nadE, and obtained the strains E. coli BW25113 (ΔnadD, NTT4) and BW25113 (ΔnadE, NTT4). Cell growth of these two strains are depending on exogenous NAD+ supplemented in the media, suggesting that we have successfully engineered E. coli to hold an NAD+ auxotrophic phenotype. We are carrying out a number of experiments using these NAD+ auxotrophic strains to address some interesting questions which may not be able to do otherwise. Results will be discussed during the conference.
英文摘要: NAD+ and its reduced form NADH are essential cofactors in biological systems. They function as cofactors in over 300 redox reactions in vivo1. The level of NAD+ and NADH is tightly controlled by a variety of mechanisms including their biosynthesis and salvage. Therefore, it is difficult to answer some fundamental questions such as the minimal NAD+ level for cell growth and the biological consequences of abnormal activity of a specific NAD+-dependent enzyme. We expressed the NTT4 gene from Chlamydiae UWE25 in Escherichia coli BW25113, as the NTT4 protein was reported as a NAD+ transporter that can specifically transport intact NAD+ across cytoplasmic membrane2. We knocked out the nadC gene responsible for de novo biosynthesis of NAD+ and constructed the strain E. coli BW25113 (ΔnadC, NTT4). It was found that NAD+ in the culture media could significantly promote the growth of BW25113 (ΔnadC, NTT4), suggesting that the NTT4 protein was functional in E. coli (Fig A, B). We then disrupted the other two genes, nadD and nadE, and obtained the strains E. coli BW25113 (ΔnadD, NTT4) and BW25113 (ΔnadE, NTT4). Cell growth of these two strains are depending on exogenous NAD+ supplemented in the media, suggesting that we have successfully engineered E. coli to hold an NAD+ auxotrophic phenotype. We are carrying out a number of experiments using these NAD+ auxotrophic strains to address some interesting questions which may not be able to do otherwise. Results will be discussed during the conference.
语种: 英语
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116038
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Zhou YJ,Hou SH,Zhang SF,et al. Construction of E. coli NAD+ auxotrophic strains and the biotechnological application thereof[C]. 见:Asian Congress on Biotechnology. 上海. 2011-5-11.
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