HIV—2gag蛋白在大肠杆菌中的表达纯化

目的 在原核系统中高效表达及纯化人Ⅱ型免疫缺陷病毒（ＨＩＶ－２）ｇａｇ蛋白，以期作为检测试剂的原料。方法 采用基因工程手段将编码Ⅱ型人免疫缺陷病毒（ＨＩＶ－２）核心蛋白的部分ｐ５５ｇａｇ１（Ｇ１）及全部ｐ５５ｇａｇ２（Ｇ２）基因片段分别克隆到原核高效表达载体ｐＥＴ２８ａ的Ｔ７噬菌体启动子下游，构建出重组表达质粒ｐＥＴ２８ａＧ１和ｐＥＴ２８ａＧ２，将其转入不同的受体菌后，ＩＰＴＧ诱导下进行表达。经Ｓ

Expression and Purification of HIV 2 gag Protein in E coli

Objective To develop a test reagent with high level expression and purification of HIV 2 gag protein in prokaryotic system Methods Expression with inserting HIV 2 p55gag2 (G2) and truncated p55gag1 (G1) into down stream of T7 promoter of prokaryotic expression vector pET28a ,induced by IPTG and analyzed with SDS PAGE and Western blot Purification with Ni NTA affinity chromatography under the conditions of both denaturation and undenaturation Results Two recombinant plasmids, pET28aG1 and pET28aG2 ,were constructed, which were highly expressed in BL21(DE3)plysS The products of pET28aG1 and pET28aG2 were 55kDa and 61kDa in weight, respectively The former is 10%and the later is 6 8%in the total protein It was shown by Western blot that the expressed recombinant protein could specifically react with HIV 2 antiserum Conclusion HIV 2 gag protein,expressed and purified in E coli, with good immunogenicity, could be used for the test reagent