Transmembrane structure and function of dctPQM encoding C4-dicarboxylate transport proteins from nitrogen-fixing *** A1501

作     者：YAN Chunling LIN Min WAN Yusong HOU Shengqiang PING Shuzhen CHEN Ming ZHANG Baoming C. Elmerich 

作者机构：Biotechnology Research Institute Chinese Academy of Sciences Beijing 100094 China Institut des Sciences du Vegetal Bâ timent 23 Avenve de la Terrasse 91198 Gif sur Yvette France 

出 版 物：《Chinese Science Bulletin》 (中国科学通报)

年 卷 期：2003年第48卷第17期

页      面：1811-1815页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071007[理学-遗传学] 071005[理学-微生物学] 10[医学] 

基　　金：supported by the National Basic Research Priorities Programme(Grant No.001CB108904) the National Natural Science Foundation of China(Grant No.39580001) 

主　　题：固氮生物 假单胞菌 C4-二羧酸 转运蛋白 dctPQM基因 核苷酸序列分析 横跨膜结构 基因编码 

摘      要：C4-dicarboxylate transport proteins of di-azotroph Pseudomonas stutzeri were encoded by dctPQM genes. Nucleotide sequence analysis indicated that dctP, dctQ, and dctM grouped together. Its nucleotide and amino acid sequence shared high homology with that of dctP gene en-coding periplasmic C4-dicarboxylate-binding protein and dctQM genes encoding C4-dicarboxylate transport proteins from the free-living nitrogen-fixing Aotobacter vinelandii. Structural analysis showed that DctP of P. stutzeri did not include membrane-spanning regions, and DctQ and DctM contained 5 and 12 transmembrane segments, respectively. The fragment containing the complete dctPQM genes was cloned into the Tn5 transposon region of suicide mobiliza-tion plasmid pSZ21. The resultant plasmid was named pSZY6. By triparental mating, Tn5 transposon carrying the dctPQM genes inserted into the genome of the wild type strain A1501, randomly. The recombinant strain A-142 which harboured an extra copy of dctPQM genes was con-structed and identified by PCR amplification of npt II gene. When A-142 was grown in minimal medium with different concentrations (20, 10 and 5 mmol/L) of C4-dicarboxylates succinate, malate, or fumarate as the sole carbon source, the rate of nitrogen fixation assayed by acetylene reduction was significantly higher than that of the wild-type strain A1501. This result was established that an extra copy of dctPQM genes could increase the activity of nitrogen fixation of P. stutzeri strain A1501.