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在鱼腥藻7120中建立反义 glnA系统

秦京东, 施定基, 徐旭东, 张金栋, 郭平仲, 汤佩松

中科院植物研究所.北京 100093；北京大学技术物理系.北京 100871；首都师范大学生物系.北京 100037；美国密执安州立大学国家植物学实验室.密执安

摘要

构建了含有反义 glnA基因片段的穿梭表达质粒,并用它转化鱼腥藻7120,获得的工程株显示了GS3受抑制和向胞外泌氨高于原始种的特性。

关键词： 反义glnA系统; 谷氨酰胶合成酶(GS); 鱼腥藻7120; 泌氨; 代谢调控

The establishment of antisense glnA system in cyanobacterium Anabaena sp. PCC7120

Qin Jingdong, Shi Dingji, Xu Xudong, Zhang Jindong, Guo Pingzhong, Tang Peisong

Institute of Botany,Chinese Academy of Sciences.Beijing 100093；China；Capital Normal University.Beijing 100037；MSU-DOE National Plant Research Lab.,Michigan State University,East Lansing,MI48824,USA

Abstract

In this work, we tried to use the antisense technique to regulate the expression of endogenic glnA gene of Anabaena sp. PCC7120, and got an strain in which antisense glnA system had been established. First, a fragment of glnA gene coding glutamine synthetase (GS) was cut down and linked with an expressional plasmid pRL-439 and a shuttle plasmid pDC-8 to get recombinants, and we screened antisense shuttle expression plasmid pDC-ATGS by endonuclease digesting. Then it was transferred into Anabaena sp. PCC7120 by triparent conjugation transfer. Neomysin screening, endonuclease digestion, dot blotting, plasmid cross transformation, observation of cell morphologic changes, and determination of activities of glutamine synthetase and NH~(+)_(4) excretion of the cells were used to confirm that the strain had an artificial antisense RNA system.

Key words: Antisense glnA system;Glutamine syntase(GS);Anabaena sp.pCC7120;Ammonia excretion;Metabolism regulation

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引用本文：

秦京东, 施定基, 徐旭东, 张金栋, 郭平仲, 汤佩松. 在鱼腥藻7120中建立反义 glnA系统[J]. 生理学报 1998; 50 (3): .

Qin Jingdong, Shi Dingji, Xu Xudong, Zhang Jindong, Guo Pingzhong, Tang Peisong. The establishment of antisense glnA system in cyanobacterium Anabaena sp. PCC7120. Acta Physiol Sin 1998; 50 (3): (in Chinese with English abstract).