Serratia sp. NDW3 GADH小亚基基因ga2dh的克隆及表达分析

发布时间：2018-03-04 09:15

本文选题：水稻　切入点：溶磷菌　出处：《华南农业大学学报》2017年02期 　论文类型：期刊论文

【摘要】：【目的】克隆葡萄糖酸脱氢酶(GADH)小亚基基因ga2dh并进行鉴定。【方法】研究水稻根际细菌Serratia sp.NDW3溶磷过程中菌株溶磷量、GADH活性与ga2dh基因表达量的变化,对ga2dh基因进行克隆和生物信息学分析,并检测ga2dh基因在大肠埃希菌BL21中的表达。【结果】Serratia sp.NDW3溶磷过程中ga2dh基因的相对表达量在12 h最大,GADH活性在24 h达到最大值,NDW3溶磷量在36 h后趋于稳定。从Serratia sp.NDW3菌株中克隆获得了781 bp的ga2dh基因序列,生物信息学分析发现该序列与Serratia sp.SCBI菌株的基因相似性为99.62%,编码的蛋白属于葡萄糖酸脱氢酶亚基3超家族,主要由3个α-螺旋构成,且氨基酸序列中包含有位于胞内、胞外和跨膜的区域。ga2dh基因在大肠埃希菌BL21体内表达,能够使得菌体GADH的活性显著增加。【结论】Serratia sp.NDW3菌株溶磷的主要机制依赖于直接氧化途径,ga2dh基因编码的小亚基不仅对GADH活性起重要作用,也是介导GADH跨膜结构的重要组成部分。
[Abstract]:[objective] to clone and identify the small subunit gene ga2dh of gluconate dehydrogenase (GADH). [methods] to study the changes of the activity of GADH and the expression of ga2dh gene during phosphorus dissolution of Serratia sp.NDW3 in rice rhizosphere bacteria. Cloning and bioinformatics analysis of ga2dh gene, The expression of ga2dh gene in Escherichia coli BL21 was detected. [results] the relative expression of ga2dh gene reached the maximum at 24 h after 12 h of Serratia sp.NDW3 phosphorolysis. After 36 h, it tended to be stable from Serratia sp.NDW3 strain. The ga2dh gene sequence of 781bp was obtained. Bioinformatics analysis showed that the gene similarity between this sequence and Serratia sp.SCBI strain was 99.62, and the encoded protein belonged to the gluconate dehydrogenase subunit 3 superfamily, which was mainly composed of three 伪 -helix, and the amino acid sequence contained in the intracellular. The extracellular and transmembrane region. Ga2dh gene was expressed in BL21 of Escherichia coli. [conclusion] the main mechanism of phosphorus solubilization of Serratia sp.NDW3 strain depends on the direct oxidation pathway. The small subunit encoded by ga2dh gene not only plays an important role in the activity of GADH, but also plays an important role in mediating the transmembrane structure of GADH.
【作者单位】：吉林农业大学生命科学学院;吉林农业大学农学院;
【分类号】：S565.101

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