嗜水气单胞菌溶血素基因克隆与序列分析  

Cloning and sequence analysis of hemolysin gene from Aeromonas hydrophila

作　　者：胡秀彩[1] 吕爱军[1,2] 孙敬锋[1] 陈成勋[1] 李莉[2] 孔祥会[2] 

机构地区：[1]天津农学院水产学院天津市水产生态及养殖重点实验室,天津300384 [2]河南师范大学水产学院,河南新乡453007

出　　处：《生物技术》2015年第6期511-514,共4页Biotechnology

基　　金：国家自然科学基金项目(No.31272692);天津市应用基础与前沿技术研究重点项目(15JCZDJC34000)资助

摘　　要：[目的]对嗜水气单胞菌溶血素基因(HlyA)进行克隆、序列分析和蛋白三级结构预测。[方法]根据嗜水气单胞菌溶血素基因(HlyA)设计特异性引物,采用PCR方法扩增并亚克隆至pMD18-T载体中,进行DNA测序分析。[结果]获得Hly A基因片段大小为1 022 bp,推导编码297个氨基酸,其中1-30aa区域为信号肽序列,7-196aa区域为溶血素-N端结构域(PF12563),222-297aa区域为杀白细胞素结构域(PF07968)。HlyA蛋白三级结构与模板(PDB:1XEZ_A)相似性达41.76%,主要由β折叠、转角构成,与拉马钱德兰图方法检测Hly A蛋白空间结构基本一致。[结论]该研究有助于理解嗜水气单胞菌Hly A基因功能及其溶血机制。[Objective]Hemolysin （HlyA） gene from Aeromonas hydrophila Zfl was amplified. [Methods] PCR method and was further analyzed by bioinformatics methods. [ Results ] The sequencing results revealed that HlyA gene was 1 022 bp long encoding a protein of 297 amino acids, which has a signal peptide located at 1 - 30 amino acids, hemolysin - N domain （PF12563 ） and leukocidin domain （PF07968）. Homology modeling was used to construct three dimensional （3 D） structure of HlyA protein with the standard model （PDB：IXEZ_A） ,which was mainly consisted of [3 -sheet and turn regions. This is consistent with the resulting protein structure prediction by using Ramachandran plot analysis. [ Conclusion] The bioinformation analyses of HlyA protein help to understand the function of ttlyA gene from the Aeromonas hydrophila and its hemolysis mechanism.

关 键 词：嗜水气单胞菌 溶血素 序列分析 三级结构 

分 类 号：Q785[生物学—分子生物学] Q751