Rolls M M, Webster P, Balba N H, Rose J K
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510.
Cell. 1994 Nov 4;79(3):497-506. doi: 10.1016/0092-8674(94)90258-5.
Self-propagating infectious particles were produced in animal cells transfected with an RNA replicon encoding a single viral structural protein, the vesicular stomatitis virus glycoprotein (VSV-G). The replicon is derived from an alphavirus, Semliki Forest virus (SFV), and encodes the SFV RNA replicase, but none of the SFV structural proteins. After transfection of the replicon into tissue culture cells, expression of G protein spread from small foci throughout the culture. Supernatants from the cells contained infectious, virus-like particles that could be passaged and were neutralized by anti-VSV serum. The majority of the infectious particles were smaller and less dense than either VSV or SFV. Characterization by electron microscopy showed membrane-enveloped vesicles that contained the VSV-G protein. Infectious particles were apparently generated by budding of vesicles containing VSV-G protein and the RNA replicon. These experiments reveal that an enveloped infectious agent can be much simpler than previously thought.
在用编码单一病毒结构蛋白——水泡性口炎病毒糖蛋白(VSV-G)的RNA复制子转染的动物细胞中产生了自我传播的感染性颗粒。该复制子来源于一种甲病毒——辛德毕斯病毒(SFV),编码SFV RNA复制酶,但不编码任何SFV结构蛋白。将复制子转染到组织培养细胞后,G蛋白的表达从小病灶扩散到整个培养物中。细胞的上清液中含有传染性的、病毒样颗粒,这些颗粒可以传代并被抗VSV血清中和。大多数感染性颗粒比VSV或SFV更小且密度更低。电子显微镜表征显示含有VSV-G蛋白的膜包裹囊泡。感染性颗粒显然是由含有VSV-G蛋白和RNA复制子的囊泡出芽产生的。这些实验表明,一种包膜感染因子可能比以前认为的要简单得多。