Fitzpatrick D R, Zamb T J, Babiuk L A
Department of Veterinary Microbiology, University of Saskatchewan, Saskatoon, Canada.
J Gen Virol. 1990 May;71 ( Pt 5):1215-9. doi: 10.1099/0022-1317-71-5-1215.
Bovine MDBK cells were transfected with Rous sarcoma virus-based vectors for constitutive expression of the bovine herpesvirus type 1 (BHV-1) glycoprotein, gI. Cell lines stably expressing recombinant gI were cloned and characterized. Recombinant gI was localized intracellularly, predominantly in a perinuclear region, and on the cell surface. Cells expressing gI exhibited spontaneous polykaryon formation, thus confirming the fusogenic activity described previously in gI-expressing transfected murine LMTK- cells. The recombinant form of gI synthesized in transfected MDBK cells was similar in Mr to the form expressed in BHV-1-infected MDBK cells, unlike the recombinant form of gI expressed by LMTK- cells which is deficient in N-linked glycosylation. It was concluded that cell fusion associated with the expression of BHV-1 gI in transfected mammalian cells is a reproducible phenomenon in a number of cell types and is not due to species-specific factors or expression of abnormally glycosylated gI. Cell fusion is a useful in vitro marker for gI function and may contribute to the spread of BHV-1 infections in vivo.
用基于劳氏肉瘤病毒的载体转染牛MDBK细胞,以组成型表达牛疱疹病毒1型(BHV-1)糖蛋白gI。对稳定表达重组gI的细胞系进行克隆和鉴定。重组gI定位于细胞内,主要位于核周区域,也存在于细胞表面。表达gI的细胞表现出自发性多核体形成,从而证实了先前在表达gI的转染鼠LMTK-细胞中描述的融合活性。转染的MDBK细胞中合成的重组gI的分子量与BHV-1感染的MDBK细胞中表达的形式相似,这与LMTK-细胞表达的缺乏N-连接糖基化的重组gI形式不同。得出的结论是,在转染的哺乳动物细胞中与BHV-1 gI表达相关的细胞融合在多种细胞类型中是一种可重复的现象,并非由于物种特异性因素或异常糖基化gI的表达所致。细胞融合是gI功能的一种有用的体外标志物,可能有助于BHV-1在体内的感染传播。