Xu J, Scalzo A A, Lyons P A, Shellam G R
Department of Microbiology, University of Western Australia, Queen Elizabeth II Medical Centre, Nedlands.
Virology. 1994 Oct;204(1):466-70. doi: 10.1006/viro.1994.1556.
A series of overlapping fragments spanning the entire coding sequence of the gH gene of murine cytomegalovirus (MCMV) were expressed in Escherichia coli as fusion proteins with glutathione S-transferase (GST) using the pGEX expression system. A region of antibody-binding was mapped to the NH2-terminus of glycoprotein H (gH) between amino acid residues 26 and 90 on the basis of the reactivity of GST-gH fusion proteins with polyclonal antibodies to MCMV in Western blot analysis. Antibodies to gH were generated in mice immunized with the GST-gH fusion protein SK and shown to react with an 87-kDa polypeptide present in virion particles which was conserved in MCMV isolates obtained from diverse locations. They also recognized the gH protein in MCMV-infected cells, as well as gH expressed in Chinese Hamster Ovary cells. The antibodies to gH had a significant ELISA titer but no neutralizing activity in vitro. The antibody response to the GST-gH fusion protein did not modify the level of MCMV replication in the spleens of mice.
利用pGEX表达系统，在大肠杆菌中表达了一系列跨越鼠巨细胞病毒（MCMV）gH基因整个编码序列的重叠片段，这些片段作为与谷胱甘肽S-转移酶（GST）的融合蛋白。基于GST-gH融合蛋白在蛋白质印迹分析中与抗MCMV多克隆抗体的反应性，将抗体结合区域定位到糖蛋白H（gH）氨基末端的26至90个氨基酸残基之间。用GST-gH融合蛋白SK免疫小鼠产生了抗gH抗体，这些抗体与病毒体颗粒中存在的一种87 kDa多肽发生反应，该多肽在从不同地点获得的MCMV分离株中是保守的。它们还识别MCMV感染细胞中的gH蛋白以及中国仓鼠卵巢细胞中表达的gH。抗gH抗体具有显著的ELISA效价，但在体外没有中和活性。对GST-gH融合蛋白的抗体反应并未改变小鼠脾脏中MCMV的复制水平。
