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_医学论文.docVIP

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_医学论文 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_医学论文 作者：冯宇, 万敏, 王宣军, 张培因, 于永利, 王丽颖 【关键词】 SARS冠状病毒刺突蛋白,B细胞表位预测,伴侣10分子,S2亚基 　　Expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and identification of its mimic antigenicity 　　　　[Abstract] AIM: To study the expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli 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 pET28chap10S2epi plasmid. The fusion protein, chap10S2epi, was expressed in E.coli BL21(DE3) and identified by SDSPAGE and Western blot. The rabbit was immunized by purified Chap10S2epi for the preparation of antiserum, which was used to identify the mimic antigenicity of Chap10S2epi to S2 protein by ELISA. RESULTS: Chap10S2epi fusion protein was successfully constructed and expressed in E.coli. The antiserum from the animal immunized by Chap10S2epi 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. 　　[Keywords]SARS spike protein B cell epitope prediction chaperone 10 S2 subunit 　　[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pET28chap10S2epi。重组蛋白Chap10S2epi在E.coli BL21(DE3)中表达并以SDSPAGE和Western blot进行鉴定。用纯化的chap10S2epi免疫家兔制备抗血清, 并通过ELISA判断Chap10S2epi的抗原性。结果: 成功构建并在大肠杆菌中表达了Chap10S2pei融合蛋白。Chap10S2epi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