重组HIV抗原和多表位抗原的研究

目的研究重组HIV病毒系列抗原,以满足HIV各种抗体检测试荆研究的需要。方法从HIV-1／2不同抗原中精选出优势抗原片段,分别用PCR方法从编码全长HIV-1基因的pBHIOR2／HIV质粒中扩增出HIV-1／gpl20、HIV-1／GP41及HIV-1／“M”优势抗原表位,用基因合成法,合成HIV-1／“0”和HIV-2／gp36基因,将获得的各基因片段插入到pBVIL1载体,使其在HB101中表达。利用载体pBVILl具有使小分子肽基因间可方便连接的特点,选择优势抗原表位进行连接和嵌合表达。结果与结论所克隆的单片段及嵌合抗原均在大肠杆菌中获得了高效的表达,纯化的单片段及多表位嵌合抗原经用间接EIA法对HIV阴性和阳性血清测定,表明抗原已满足抗体检测试剂盒的要求。得到的单片段抗原可用于LIA测定,而多表位嵌合抗原,可用于EIA筛检试剂。

Study on Recombinant HIV Antigens and Multi-epitope Antigen

Objective:To study the recombinant HIV antigens from different HIV proteins to fit the need in different kinds of antibody assays.Methods A series of HIV-1 and HIV-2 antigen fragments were selected from different proteins expressed by the virus genes.The genes of these antigens were acquired from a plasmid, pBH1OR2/HIV containing the whole gene of HIV-1 by PCR.For the gene of HIV-2 and HIV-1/"0" immunodominant region(IDR) were chemically synthesized directly.All these genes were cloned into an expresion plasmid pBVIL1 prepared in our laboratory.And owing to the character of pBVIL1,a chimera gene containing HIV-1/gp41,HIV-2/gp36 and HIV-1/"0" IDR was constructed and expressed in E.coli too.Results All the genes including the chimera cloned above were highly expressed in E.coli and all the purified recombinant peptides were specifically reacted with HIV positive sera.Conclusion These results suggest that the recombinant IDR peptides can be used in LIA assay and the chimera will be good at EIA screen assay.