框镜鲤维氏气单胞菌CY0806株鞭毛flaA基因的克隆与生物信息学分析

Cloning and Bioinformatic Analysis of Flagellum flaA Gene of Aeromonas veronii CY0806 from Cyprinus carpio

以框镜鲤Cyprinus carpio维氏气单胞菌Aeromonas veronii CY0806株细菌基因组DNA为模板,经PCR扩增,获得flaA基因,将其克隆并测序,测序结果与气单胞菌属的不同菌株进行相似性分析,并对flaA基因编码蛋白质进行生物信息学分析.结果显示:获得框镜鲤维氏气单胞菌鞭毛flaA基因,长915 bp,编码304个氨基酸,系统进化树分析,其与其他维氏气单胞菌flaA基因处于同一分支,相关生物信息学分析显示,flaA基因编码的蛋白质相对分子质量32 098.66,等电点5.57,半衰期30 h,不稳定系数为32.31,脂肪系数81.05,不存在信号肽和跨膜区,二级结构预测:flaA基因以α螺旋和β折叠为主,同时,以同源建模法预测了flaA基因编码蛋白的三维立体结构.

The flaA gene was amplified from genome template of Aeromonas veronii CY0806 of Cyprinus carpio by PCR. Homogeneous analysis was carried out by comparing the sequencing results with other strains of Aeromonas and bioinformatic analysis was done on the protein coded by flaA. The results showed that flaA Gene with a length of 915 bp, coding 304 amino acids, was obtained ; phylogenetic tree analysis results showed that it was on the same branch with other strains of Aeromonas. Bioinformatic analysis results showed that the relative molecular mass of the protein was 32 098.66, pI 5.57, half-life 30 h, unstability index 32. 31, fat index 81.05 whereas there was no signal sequence and transmembrane domain. Results of secondary structural analysis showed that the main secondary structures were α-helix and β-sheet. At the same time the tertiary structure was also predicted by homology modeling.