细粒棘球绦虫免疫相关抗原基因的筛选及原核表达  

Screening of immune-related antigenic genes of Echinococcus granulosus and the prokaryotic expression

作　　者：刘原源 安红燕[1] 杨爽[1] 王晓岑[1] 宫鹏涛[1] 张壮志[2] 李建华[1] 杨举[1] 李赫[1] 张西臣[1] 

机构地区：[1]吉林大学动物医学学院,长春130062 [2]新疆畜牧科学院兽医研究所,乌鲁木齐830000

出　　处：《黑龙江畜牧兽医》2016年第11期149-151,155,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家公益性行业(农业)科研专项(201303042)

摘　　要：为了筛选获得细粒棘球绦虫的免疫相关抗原基因,试验采用SEREX技术对细粒棘球绦虫cDNA文库进行相关抗原筛选,并对获得的细粒棘球绦虫特异性基因进行原核表达与鉴定。结果表明:共获得149个阳性噬菌斑,对其进行PCR扩增和序列分析得到与棘球绦虫属相关的基因序列10个,其中有864 bp的开放性阅读框序列;编码288个氨基酸;经NCBI BLAST氨基酸同源性分析,与多房棘球绦虫诊断抗原gp50的氨基酸序列的同源性为92%,是一个未知的新基因,命名为Ediag A864;经原核表达得到大小约为51 ku的重组蛋白,经Western-blot分析具有良好的反应原性。To screen and obtain immune -related antigenic genes of Echinococcus granu/osus,SEREX technology was used to screen the related antigens from a eDNA library of Echiaococcus grantdosus, and then a specific gene obtained from Echinococcus grantdosus was used for prokary- otic expression and identification. The result showed that a total of 149 positive plaques were obtained, and 10 gene sequences associated with the Echinococcus genus were obtained by PCR amplification and sequence analysis. A specific gene was obtained, which had an open reading frame of 864 bp, encoding for a protein of 288 amino acids. The homology analysis by NCBI BLAST revealed that the amino acid sequence of the gene shared 92% homology with that of E. multilocularis diagnostic antigen gp50. The gene was an unknown new gene, which was named EdiagA864. A recombinant protein with a size of about 51 ku was obtained by prekaryotic expression and the protein had a good reactianogenicity by Western -blot.

关 键 词：细粒棘球绦虫 cDNA文库 基因筛选 原核表达 免疫相关 同源性分析 

分 类 号：S852.734[农业科学—基础兽医学] Q786[农业科学—兽医学]