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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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Citing This Article：

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).