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

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.