| 外源glnA基因干扰大肠杆菌的代谢 |
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| 引用本文： | 杜娟,吴健,戴桂馥,王春阳,周新钦,宋明辉,李珏,李季伦.外源glnA基因干扰大肠杆菌的代谢[J].生物工程学报,2009,25(4):626-631. |
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| 作者姓名： | 杜娟 吴健 戴桂馥 王春阳 周新钦 宋明辉 李珏 李季伦 |
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| 作者单位： | 郑州大学生物工程系,郑州,450001 |
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| 基金项目： | 河南省重点科技攻关项目(No.0624240002)资助~~ |
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| 摘 要： | 采用分子生物学的方法构建了含Bacillus subtilis glnA基因的重组菌株Escherichia coli DH5α(pMD19-glnA),用毛细管电泳和核磁共振对重组菌株的转化谷氨酸的产物进行定性鉴定,并进一步通过荧光定量RT-PCR测定谷氨酰胺合成酶基因(glnA)mRNA水平的相对表达量,最后用SDS-聚丙烯酰胺凝胶电泳对蛋白的相对表达情况进行了分析。结果表明重组菌株并没有增加谷氨酰胺的产量,而是明显增加了γ-氨基丁酸(GABA)的产量。实验表明重组菌株中的glnA基因可以正常转录,但是谷氨酰胺合成酶的蛋白表达量并没有增加。这种外源基因干扰大肠杆菌代谢的现象值得进一步研究。
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| 关 键 词： | 谷氨酰胺合成酶 毛细管电泳 荧光定量RT-PCR SDS-聚丙烯酰胺凝胶电泳 |
| 收稿时间： | 2008/9/27 0:00:00 |
Metabolism of Escherichia coli is interfered by Bacillus subtilis glnA gene |
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| Juan Du,Jian Wu,Guifu Dai,Chunyang Wang,Xinqin Zhou,Minghui Song,Jue Li and Jilun Li.Metabolism of Escherichia coli is interfered by Bacillus subtilis glnA gene[J].Chinese Journal of Biotechnology,2009,25(4):626-631. |
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| Authors: | Juan Du Jian Wu Guifu Dai Chunyang Wang Xinqin Zhou Minghui Song Jue Li and Jilun Li |
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| Institution: | Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China;Department of Biotechnology, Zhengzhou University, Zhengzhou 450001, China |
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| Abstract: | A recombinant strain Escherichia coli DH5a(pMD19-glnA) including Bacillus subtilis glnA gene was constructed. Capillary electrophoresis and nuclear magnetic resonance were used to determine qualitatively the product of transformation by recombinant strain, and the relative level of mRNA expression of glnA was also determined by fluorescence quantitative RT-PCR. Subsequently, SDS-PAGE (polyacrylamide gel electrophoresis) was used to analysis the relative level of protein. Surprisingly, there was no increase of glutamine production in this recombinant strain, but an obvious increase in the GABA (g-aminobutyric acid ) production. It was showed in the experiment that protein expression of the glutamine synthetase did not increase, although glnA gene can be transcribed normally in this recombined strain. The phenomenon of exogenous glnA gene interfering metabolism of Escherichia coli was worthy of further study. |
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| Keywords: | glutamine synthetase (GS) capillary electrophoresis fluorescence quantitative RT-PCR SDS-PAGE |
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