Cloning and Identification of Ammonia Monooxygenase Gene of Sinorhizobium sp. NP1

LIU Yu-Ying,QIU Feng,SONG Qin,DOU Xin,XU Lei

Chinese Journal of Biochemistry and Molecular Biology ›› 2010, Vol. 26 ›› Issue (8) : 768-775.

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Chinese Journal of Biochemistry and Molecular Biology ›› 2010, Vol. 26 ›› Issue (8) : 768-775.
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Cloning and Identification of Ammonia Monooxygenase Gene of Sinorhizobium sp. NP1

  • LIU Yu-Ying 1) ,QIU Feng 2) ,SONG Qin 1) ,DOU Xin 1) ,XU Lei 1)*
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Abstract

The ammonia monooxygenase (AMO) gene of 1 089 bp in Sinorhizobium sp. NP1, a strain established by our laboratory, was cloned by homology method and Tail-PCR. The gene encodes 362 amino acids and the predicted secondary structure with nine transmembrane regions is similar to that of Sinorhizobium meliloti. We have constructed a NP1 amo knockout plasmid named pJQ200SK-amo-Tc and transformed into the original NP1 strain using three parent hybridization and obtained a amo knockdown strain. The enzymatic activities in NP1∶∶ amo decreases about 35% compared to original NP1 strains by Berthelot test. The results showed that amo encoded a key enzyme for denitrification.

Key words

ammonia monooxygenase (AMO) / gene cloning / gene knockout / biological nitrogen removal

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LIU Yu-Ying,QIU Feng,SONG Qin,DOU Xin,XU Lei. Cloning and Identification of Ammonia Monooxygenase Gene of Sinorhizobium sp. NP1[J]. Chinese Journal of Biochemistry and Molecular Biology, 2010, 26(8): 768-775

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Funding

Supported by National High Technology Research and Development Program of China (863 Program, No.2006AA10C413)

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