Lyles D S, Puddington L, McCreedy B J
Department of Microbiology and Immunology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103.
J Virol. 1988 Nov;62(11):4387-92. doi: 10.1128/JVI.62.11.4387-4392.1988.
The M protein of vesicular stomatitis virus (VSV) was localized in the nuclei and cytoplasm of VSV-infected cells by subcellular fractionation and immunofluorescence microscopy. Nuclei isolated from VSV-infected Friend erythroleukemia cells were fractionated into a nuclear membrane and a nucleoplasm fraction by DNase digestion and differential centrifugation. G protein was present in the membrane fraction, and M protein was present in the nucleoplasm fraction. Immunofluorescence detection of M protein in the nucleus required that fixed cells be permeabilized with higher concentrations of detergent than were required for detection of M protein in the cytoplasm of VSV-infected BHK cells.
通过亚细胞分级分离和免疫荧光显微镜技术,水泡性口炎病毒(VSV)的M蛋白定位于VSV感染细胞的细胞核和细胞质中。从VSV感染的Friend红白血病细胞中分离出的细胞核,通过DNA酶消化和差速离心被分级分离为核膜部分和核质部分。G蛋白存在于膜部分,而M蛋白存在于核质部分。在细胞核中对M蛋白进行免疫荧光检测时,与检测VSV感染的BHK细胞细胞质中的M蛋白相比,需要用更高浓度的去污剂使固定细胞通透化。