HIV—1长末端重复序列对重组痘苗病毒表达Gag蛋白的影响

将系列缺失的ＨＩＶ１长末端重复序列（ＬＴＲ）和全长的ｇａｇＯＲＦ置于痘苗病毒载体中，经同源重组和血球吸附试验，成功地构建了６株重组痘苗病毒。免疫印迹和免疫酶试验检测均表明，６株重组病毒的Ｇａｇ蛋白表达量因ＬＴＲ不同而有明显差异，表明ＨＩＶ１的ＬＴＲ及其下游基因置于痘病毒启动子控制下，在痘苗病毒中表达时有下述特点：（１）不同的痘苗病毒启动子与全长ＬＴＲ相互作用，对ｇａｇ基因表达有显著不同的调控效果；（２）ＮＲ序列对Ｇａｇ蛋白表达没有明显影响；（３）ＥＮ序列不能被重组痘苗病毒表达系统识别；（４）ＴＡＲ序列可提高Ｇａｇ蛋白的表达量；（５）Ｕ５区及下游非翻译序列不影响Ｇａｇ蛋白的表达。

Effects of Long Terminal Repeat (LTR) of Human Immumodeficiency Virus Type 1 on the Expression of gag Gene in Recombinant Vaccinia Viruses

To investigate the function of LTR in vaccinia virus, 6 recombinant viruses were constructed by inserting intact or serially truncated LTR together with their downstream of gag ORF of human immunodeficiency virus type 1 (HIV 1) into the vaccinia virus genome. Indirect fluorescence assay, Western blot and immunoenzyme assay showed that all the 6 recombinant vaccinia viruses expressed Gag protein, but the efficiency were significantly different. These results suggested that the function of each function domain in HIV 1 LTR exhibited distinct characteristics on gag gene expression in vaccinia virus: (1) The function of intact LTR on gag gene expression varied with the upstream poxvirus promoter; (2) NR sequence neither decreased nor increased the Gag protein exprssion level; (3) Enhancer sequence might not be recognized by recombinant vaccinia virus expression system; (4) TAR sequence enhanced Gag protein expression effectively; (5) U5 region and its downstream non translated sequence had little effect on Gag protein exprssion efficiency.