Fang B, Xu B, Koch P, Roth J A
Department of Thoracic and Cardiovascular Surgery, University of Texas MD Anderson Cancer Center, Houston 77030, USA.
Gene Ther. 1998 Oct;5(10):1420-4. doi: 10.1038/sj.gt.3300741.
Herpes simplex virus type 1 (HSV-1) VP22 was recently reported to mediate intercellular trafficking of a protein fused to the C-terminus of VP22. To explore the application of such trafficking, we constructed plasmids expressing green fluorescent protein (GFP) fused to the C-terminus of either wild-type VP22 or a 160 amino acid peptide from VP22. In vitro studies showed that the majority of both fused proteins were localized to the nuclei of transfected cells. Quantitative analysis of GFP-positive cells, however, showed no significant increase in intercellular protein trafficking for cells transfected with either fusion protein compared with a lacZ-expressing plasmid. Our results suggest that the use of HSV-1 VP22 for mediating intercellular trafficking of transgene products is limited.
最近有报道称,单纯疱疹病毒1型(HSV-1)的VP22可介导与VP22 C末端融合的蛋白质的细胞间转运。为了探索这种转运的应用,我们构建了表达与野生型VP22或来自VP22的160个氨基酸肽的C末端融合的绿色荧光蛋白(GFP)的质粒。体外研究表明,两种融合蛋白的大部分都定位于转染细胞的细胞核中。然而,对GFP阳性细胞的定量分析表明,与表达lacZ的质粒相比,用任何一种融合蛋白转染的细胞在细胞间蛋白质转运方面均未显著增加。我们的结果表明,使用HSV-1 VP22介导转基因产物的细胞间转运是有限的。