人免疫缺陷病毒1型vpu基因在大肠杆菌中的表达

Expression of Human Immunodeficiency Virus Type 1 vpu Gene in E coli

将编码人１型免疫缺陷病毒（ＨＩＶ－１）蛋白Ｕ的ｖｐｕ基因片段，克隆到原核表达载体ｐＥＴ－１７ｂ２（ｐＥＴ－１７ｂ去掉了一含起始密码子ＡＴＧ的６０ｂｐ小片段）的Ｔ７噬菌体启动子下游，构建成重组表达质粒ｐＥＴ－１７ｂ２，并使ｖｐｕ基因片段在大肠杆菌ＢＬ２１（ＤＥ３）中高效表达，产物为１６０００的Ｖｐｕ重组蛋白，表达量占菌体总蛋白的１１．２％。蛋白质印迹试验表明。

An expressive plasmid pET 17b2was constructed by inserting the vpu gene of human immunodeficiency virus type1(HIV 1)into the downstream of the bacteriophage T7promoter.Itwas shown by SDS PAGE that a16000recombinant protein was successfully expressed with ahigh level of approximately11.2%of the total bacterial protein.Western blot exhibited that the recombi nant protein could be recognized by anti vpu monoclonal antibody.