摘要
为制备高纯度p24 抗原,在变性条件下,将大肠杆菌表达的重组p24 蛋白,使用Ni-NTA 亲和层析法进行了纯化。对包涵体中的p24 蛋白进行层析纯化时,回收率略有提高,但蛋白纯度(81% )低于菌体裂解纯化洗脱物(94% )。菌体中及纯化、复性后的目的蛋白均能与抗HIV-1 p24 单克隆抗体发生特异性反应。用纯化的p24 蛋白免疫小鼠,4 周时小鼠血清抗p24 平均抗体效价达1∶400。试验结果表明,大肠杆菌表达的HIV-1 p24 全菌体裂解物纯化后,可用作HIV-1 检测试剂的原料。
A procedure is described here that allows the rapid purification of the HIV 1 p24 protein fused to both amino and carboxy terminal tag of six histidines. Extracts from the induced E.coli were loaded onto Ni 2+ nitrilotriacetic acid (Ni 2+ NTA) agarose and histidine tagged proteins were selectively eluted to >94% purity under denaturation condition. The purified p24 gag protein could be recognized by anti p24 antibody in western blot, and was capable of priming mice for the humoral response to HIV 1 Gag protein expressed in the recombinant vaccinia viruses. This part of experiment promises to be useful for the preparation of p24 gag specific antibody and diagnostic material.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1999年第6期519-522,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金
国家杰出青年基金
