HIV－1外膜（env）基因在重组痘苗病毒中的高效表达

HIGH LEVEL EXPRESSION OF HIV-1 ENV PROTEIN IN RECOMBINANT VACCINIA VIRUS

应用重组痘苗病毒作载体，在ＡＴＩ、Ｐ７．５突变型早期启动子控制下，高效表达了（经定点突变去除一处早期终止信号的）ＨＩＶ－１ｅｎｖ基因。经Ｗｅｓｔｅｒｎｂｌｏｔ证明，晚期启动子活性强的ｐＳＦＪ２－３８能更高效地表达ｅｎｖ基因。经Ｌｅｎｔｉｌ－Ｌｅｃｔｉｎ提取以及ＳＤＳ－ＰＡＧＥ后的激光扫描测定结果，在１０７个感染细胞中可产生５０－７０μｇ的Ｅｎｖ蛋白。

recombinant vaccinia virus was used as vector.Under control of ATI and P7.5 mutant early promotor,the HIV-I env gene,deprived of an early terminal signal,was successfullyexpressed to high level.The result of Western blot showed that the late promoter with highactivity,pSFJ2-38,induced higher expression of env gene.After preliminary purification with Lentil-Lectin and SDS-PAGE treatment,estimation with laser scanning technique indicated that 50-70μg of Env protein could be produced in 107 infected cells,the highest production of HIV-I Env protein in cell cultures ever reported.