Stable expression of primary human immunodeficiency virus type 1 structural gene products by use of a noncytopathic sindbis virus vector.
Stable expression of primary human immunodeficiency virus type 1 structural gene products by use of a noncytopathic sindbis virus vector.
复制标题
使用非细胞病变的辛德比斯病毒载体稳定表达原代人类免疫缺陷病毒 1 型结构基因产物。
DOI：
10.1128/jvi.76.22.11434-11439.2002
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发表时间：
2002
影响因子：
5.4
通讯作者：
Yu,Xiao-Fang
中科院分区：
文献类型：
--
作者：
Kong,Wei;Tian,Chunjuan;Liu,Bindong;Yu,Xiao-Fang
人类免疫缺陷病毒1型（HIV-1）结构基因产物Gag、Pol和Env的有效表达涉及病毒Rev和Rev应答元件（RRE）的调控。研究发现，在没有Rev和RRE的情况下，去除这些结构基因编码区的多个抑制序列（INS）或将HIV-1基因的密码子使用模式修改为高表达的人类基因所使用的密码子使用模式，可以显著增加HIV-1结构蛋白的表达。在这项研究中，我们证明了HIV-1结构基因产物Gag和Env可以在未经任何序列修饰的情况下，用无细胞病变的Sindbis病毒表达载体转染HIV-1蛋白。观察到这些Gag和Env蛋白的稳定表达超过12个月。Sindbis病毒表达载体仅在转染细胞的细胞质中复制其RNA，并且含有未经修饰的HIV-1 Gag序列的DNA载体缺乏HIV-1 Gag的表达与缺乏可检测的细胞质micgagrna相关，这一事实表明，在没有Rev/RRE的情况下，HIV-1结构蛋白表达的主要阻断是由mRNA在细胞质中的低效积累引起的。通过非细胞病变Sindbis病毒表达系统高效、长期地表达多种HIV-1毒株的结构蛋白，可能为HIV-1基因产物的功能研究和疫苗研究提供有用的工具。
Efficient expression of the human immunodeficiency virus type 1 (HIV-1) structural gene products Gag, Pol, and Env involves the regulation by viral Rev and Rev-responsive elements (RRE). Removal of multiple inhibitory sequences (INS) in the coding regions of these structural genes or modification of the codon usage patterns of HIV-1 genes to those used by highly expressed human genes has been found to significantly increase HIV-1 structural protein expression in the absence of Rev and RRE. In this study, we show that efficient and stable expression of the HIV-1 structural gene products Gag and Env could be achieved by transfection with a noncytopathic Sindbis virus expression vector by using HIV-1 sequences from primary isolates without any sequence modification. Stable expression of these Gag and Env proteins was observed for more than 12 months. The fact that the Sindbis virus expression vector replicates its RNA only in the cytoplasm of the transfected cells and the fact that the lack of expression of HIV-1 Gag by the DNA vector containing unmodified HIV-1gagsequences was associated with a lack of detectable cytoplasmicgagRNA suggest that a major blockage in the expression of HIV-1 structural proteins in the absence of Rev/RRE is caused by inefficient accumulation of mRNA in the cytoplasm. Efficient long-term expression of structural proteins of diverse HIV-1 strains by the noncytopathic Sindbis virus expression system may be a useful tool for functional study of HIV-1 gene products and vaccine research.