大肠杆菌氮代谢调节蛋白GlnG与固氮施氏假单胞菌氮代谢调节蛋白NtrC的功能互补研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.04.025

生物技术通报 ›› 2016, Vol. 32 ›› Issue (4): 190-197.doi: 10.13560/j.cnki.biotech.bull.1985.2016.04.025

大肠杆菌氮代谢调节蛋白GlnG与固氮施氏假单胞菌氮代谢调节蛋白NtrC的功能互补研究

柯琪¹，²,陈清华²,战嵛华²,陆伟²,燕永亮²

1.西南科技大学生命科学与工程学院，绵阳 621010;
2.中国农业科学院生物技术研究所，北京 100081

收稿日期:2015-12-17 出版日期:2016-04-25 发布日期:2016-04-26
作者简介:柯琪，女，硕士，研究方向：固氮微生物与基因工程，E-mail：543274382@qq.com；陈清华为本文并列第一作者
基金资助:
国家“973”项目（2015CB755700），国家自然科学基金项目（31470174），广东省引进创新创业团队计划项目（2013S033）

Functional Complement Between General Nitrogen Regulator GlnG in Escherichia coli and General Nitrogen Regulator NtrC in Pseudomonas

KE Qi¹，² ,CHEN Qing-hua², ZHAN Yu-hua², LU Wei², YAN Yong-liang²

1. School of Life Science and Engineering，Southwest University of Science and Technology，Mianyang 621010;
2. Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081

Received:2015-12-17 Published:2016-04-25 Online:2016-04-26

摘要/Abstract

摘要： 旨在围绕大肠杆菌DH10B（Escherichia coli DH10B）中氮代谢调节蛋白GlnG与施氏假单胞菌A1501（Pseudomonas stutzeri A1501）中氮代谢调节蛋白NtrC在一般氮代谢调控网络中是否存在功能互补展开研究。利用DH10B菌的glnG基因回补A1501菌的ntrC突变株，以及A1501菌的ntrC基因回补DH10B菌的glnG突变株，对所获得的功能互补株分别展开生理生化表型测定。结果表明，在以硝酸钾或尿素为唯一氮源的培养条件下，DH10B glnG基因可以回补A1501 ntrC突变株的氮源利用能力，并且部分恢复ntrC突变株的固氮酶活性（约为野生型A1501固氮酶活性的45%）；与DH10B glnG突变株相比，A1501菌的ntrC基因回补了DH10B glnG突变株以精氨酸为唯一氮源的生长能力。以上结果说明DH10B菌的GlnG蛋白与A1501菌的NtrC蛋白不仅在进化关系上紧密联系而且在所测试的氮代谢途径中存在功能互补。

关键词: 施氏假单胞菌A1501 NtrC, 大肠杆菌DH10B GlnG, 一般氮代谢, 功能互补

Abstract: This study aims to investigate whether or not there is functional complement in general nitrogen-regulation network between nitrogen metabolism regulator GlnG from Escherichia coli DH10B and NtrC from Pseudomonas stutzeri A1501. Using gene glnG of DH10B to complement gene ntrC of A1501 or vice versa，the physiological and biochemical phenotypes of the obtained functionally complementary strains were measured. The results showed that while nitrate or urea as the sole nitrogen source，DH10B’s glnG complemented the nitrogen-utilizing ability and partially restored the nitrogenase activity（about 45% of wild type）of mutant A1501’s ntrC. Comparing with DH10B’s glnG，A1501’s ntrC only complemented mutant DH10B glnG while arginine as sole nitrogen source. Above results revealed that not only was there close phylogenetical relationship，but also the functional complement in the tested nitrogen metabolism between DH10B’s GlnG and A1501’s NtrC.

Key words: Pseudomonas stutzeri A1501 NtrC, Escherichia coli DH10B GlnG, general nitrogen metabolism, functional complement

柯琪,陈清华,战嵛华,陆伟,燕永亮. 大肠杆菌氮代谢调节蛋白GlnG与固氮施氏假单胞菌氮代谢调节蛋白NtrC的功能互补研究[J]. 生物技术通报, 2016, 32(4): 190-197.

KE Qi， ,CHEN Qing-hua, ZHAN Yu-hua, LU Wei, YAN Yong-liang. Functional Complement Between General Nitrogen Regulator GlnG in Escherichia coli and General Nitrogen Regulator NtrC in Pseudomonas[J]. Biotechnology Bulletin, 2016, 32(4): 190-197.

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大肠杆菌氮代谢调节蛋白GlnG与固氮施氏假单胞菌氮代谢调节蛋白NtrC的功能互补研究

Functional Complement Between General Nitrogen Regulator GlnG in Escherichia coli and General Nitrogen Regulator NtrC in Pseudomonas

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