摘要
为深入研究人Ⅰ型免疫缺陷病毒(HIV—1)衣壳蛋白p24的结构与功能,制备抗P24单克隆抗体及其诊断抗原,将编码HIV-1p24蛋白的p24gag基因片段,克隆到原核表达载体pET-17b的T7噬菌体启动子下游,构建成了重组表达质粒pET24,并使p24gag基因片段在大肠杆菌BL21(DE3)中高效表达,其产物为30kDa的s10-p24融合蛋白,表达量占菌体总蛋白的38.4%。重组P24蛋白均抗p24单克隆抗体及HIV—1阳性血清发生特异性反应,具有较好的抗原性,对研究开发AIDS诊断试剂和基因工程疫苗具有重要意义。
In order to study the capsid protein (p24) of human immunodeficiency virus type 1(HIV-1),a plasmid vector pET24,containing the p24gaq gene fragment encoding the p24 protein under the control of the bacteriophage T7 promoter ,was constructed.The pET24 plasmidhighly expressed a 30kDa protein of s10-p24 spanning 284 amino acid residues.The total amount of the fusion protein was approximately 38.4% of the total celluar protein in Escherchia coli BL21(DE3).Westein blotting and,immunodot blotting indicated that the recombinain protein could be relognized by antip24 monoclonal antibody and HIV-1 positive sera respectively.It is significance for preparing specific antibody,and diagnostic usage.
出处
《中国人兽共患病杂志》
CSCD
北大核心
1999年第4期13-16,共4页
Chinese Journal of Zoonoses
基金
国家自然科学基金
国家杰出青年基金