预测的SARS冠状病毒刺突蛋白S2亚基B细胞表位肽在大肠杆菌中的表达及其抗原性的鉴定  

Expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and identification of its mimic antigenicity

作　　者：冯宇[1] 万敏[1] 王宣军[2] 张培因[1] 于永利[2] 王丽颖[1] 

机构地区：[1]吉林大学基础医学院分子生物学教研室,吉林长春130021 [2]吉林大学基础医学院免疫学教研室,吉林长春130021

出　　处：《细胞与分子免疫学杂志》2007年第2期113-116,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金海外杰出青年基金项目(30328010)

摘　　要：目的研究预测的SARS冠状病毒刺突蛋白S2亚基B细胞表位肽在大肠杆菌中的表达及其模拟S2蛋白的抗原性。方法用DNAStar软件对S2亚基序列进行分析,预测B细胞表位所在肽段。通过4轮PCR反应人工搭建预测表位cDNA序列并克隆到含有伴侣10基因的pET28a(+)载体中,构成重组载体pET28-chap10-S2epi。重组蛋白Chap10-S2epi在E.coliBL21(DE3)中表达并以SDS-PAGE和Westernblot进行鉴定。用纯化的chap10-S2epi免疫家兔制备抗血清,并通过ELISA判断Chap10-S2epi的抗原性。结果成功构建并在大肠杆菌中表达了Chap10-S2pei融合蛋白。Chap10-S2epi免疫的抗血清能识别真核细胞表达的SARS冠状病毒全长S2刺突蛋白。结论预测的SARS冠状病毒S2刺突蛋白B细胞表位肽能够诱导家兔产生针对S2蛋白的抗体,为研制抗SARS病毒基因工程疫苗奠定了基础。AIM： To study the expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E. coil and its mimic antigenicity to S2 protein. METHODS： B cell epitopes in S2 subunit of SARS coronavirus spike protein was predicted using DNAStar software. The cDNA sequence encoding the B cell epitope peptide was constructed artificially by PCR and then cloned into the downstream of chaperone 10 gene in vector pET28a（ ＋ ） to construct pET28-chap10-S2epi plasmid. The fusion protein, chap10-S2epi, was expressed in E. coil BL2I （DE3） and identified by SDS-PAGE and Western blot. The rabbit was immunized by purified Chap10-S2epi for the preparation of antiserum, which was used to identify the mimic antigenicity of Chap10-S2epi to S2 protein by ELISA. RESULTS： Chap10-S2epi fusion protein was successfully constructed and expressed in E. coli. The antiserum from the animal immunized by Chap10-S2epi recognized full length of SARS coronavirus S2 spike protein. CONCLUSION： The predicted B cell epitope peptide of SARS coronavirus S2 spike protein can induce the antigenicity of S2 protein, which provides some fundamental data for developing engineering vaccine against SARS coronavirus infection.

关 键 词：SARS冠状病毒剌突蛋白 B细胞表位预测 伴侣10分子 S2亚基 

分 类 号：R373[医药卫生—病原生物学]