摘要
为研究维氏气单胞菌(Aeromonas veronii)主要黏附素Aha1基因的生物信息学特性,本试验克隆了A.veronii TH0426主要黏附素Aha1基因片段,构建了Aha1基因序列进化树,通过生物信息学软件分析其编码蛋白的理化性质、疏水性、信号肽、跨膜区、二级结构及三级结构,并进行了Aha1蛋白的原核表达及免疫原性的检测。结果显示:Aha1基因全长1 038bp,编码345个氨基酸。TH0426株Aha1与A.veronii属同一分支,具有信号肽,跨膜区,二级结构以β折叠、无规则卷曲为主。SDS-PAGE及Western blot分析结果显示,Aha1蛋白在大肠杆菌中成功表达且其能被鼠抗黏附素阳性血清识别,表明Aha1蛋白具有一定的反应原性。以上结果为进一步研究Aha1蛋白的结构功能Aha1基因工程亚单位疫苗的制备奠定了基础。
The Ahal gene of A. veronii TH0426 was cloned hy PCR and sequenced, then bioinfor- matic analysis was conducted to investigate the structure and function of the Ahal gene. The phys- ical and chemical properties, hydrophobicity, the signal peptide, transmembrane region, secondary structure and tertiary structure of Ahal that the gene encoded were analyzed and predicted by using bioinformatics software,then prokaryotic expression and immunogenicity analysis of recombinant protein were carried out. The results showed that the length of Ahal gene was 1038 bp,encoding 490 amino acid residues. The Ahal gene was clustered into the same branch with A. veronii. The Ahal contained the signal peptide and transmembrane region. Secondary structure of Ahal was mainly 13-fold and random coil. SDS-PAGE and Western blot analysis show that Ahal gene was expressed in E. coli, which could be recognized by anti-Ahal mouse positive sera, indicating that the recombinant protein has a strong immunogenicity. It laid a foundation for further study of outer membrane protein genetic engineering vaccine.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第11期2141-2146,2150,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31201927)
吉林省重点科技攻关项目(20150204065NY)
作者简介
通讯作者，E-mail：qianaidong01156163．com
通讯作者，E—mail：sxfl997@163．com；
