猪内源性反转录病毒囊膜蛋白基因的克隆及其蛋白二级结构和B细胞表位预测

Clone and Prediction of the Secondary Structure and B Cell Epitopes for the Env Protein of Porcine Endogenous Retrovirus

猪内源性反转录病毒(PERV)是与猪-人异种移植病原安全性密切相关的一类病毒。env基因编码病毒的囊膜蛋白,它与病毒的亚型分类、宿主感染范围、细胞的嗜性以及对宿主细胞的感染机制、诱导宿主产生中和抗体等密切相关。本研究利用RT-PCR的方法,从五指山小型猪外周血淋巴细胞中扩增PERV的囊膜蛋白基因并进行测序,随后用生物信息学相关软件和方法,对PERV-Env蛋白二级结构及B细胞表位进行预测。经综合分析评价,结果发现PERV-Env蛋白有18个可能的B细胞优势抗原表位区域,7个可能的糖基化位点。该分析预测结果不但有利于PERV疫苗的设计、单抗及诊断试剂研制,而且将有助于分析Env蛋白的功能及PERV对人源细胞的感染机制。

Porcine endogenous retrovirus（PERV）is a kind of virus correlatively to the safety of pathogens during pig xenografts. The env gene encodes the Env protein, which plays an important role in virus subtyping, host ranging, cells tropism and mechanism of virus infection, as well as induction of neutralizing antibody secretion. In this research, the envelope gene of PERV was amplified by RT-PCR and sequenced from the peripheral blood lymphocyte of WZS minipigs, The secondary structure and B cell epitopes of the Env protein of PERV were analysed and predicted subsequently by the Bioinformatics software. The results showed that the PERV-Env protein was supposed to exist 18 potential antigen epitopes and 7 potential glycosylated sites. These results indicated that it would be helpful not only to the PERV vaccine designing and the preparation of both of monoclonal antibody and diagnostic reagent, but also to research of Env protein function and mechanism of PERV infecting human-derived cells.