重组HIV抗原和多表位抗原的研究

Study on Recombinant HIV Antigens and Multi-epitope Antigen

目的 研究重组 HIV病毒系列抗原 ,以满足 HIV各种抗体检测试剂研究的需要。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 p BH1 OR2 /HIV质粒中扩增出 HIV- 1 /gp1 2 0、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 p BVIL1载体 ,使其在 HB1 0 1中表达。利用载体 p BVIL1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

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.