Xu J, Scalzo A A, Lyons P A, Farrell H E, Rawlinson W D, Shellam G R
Department of Microbiology, University of Western Australia, Queen Elizabeth II Medical Centre, Nedlands.
J Gen Virol. 1994 Nov;75 ( Pt 11):3235-40. doi: 10.1099/0022-1317-75-11-3235.
DNA sequence analysis of the genome of the Smith strain of murine cytomegalovirus (MCMV) revealed an open reading frame (ORF) with amino acid sequence identity to glycoprotein L (gL) of other herpesviruses. The ORF is 822 bp in size and has the capacity to encode a protein of 274 amino acids. It has significant identity with the gL genes of human CMV and human herpesvirus 6. The coding sequence of the gL gene of MCMV strain K181 was also determined, and expressed in Escherichia coli as a fusion protein with glutathione S-transferase using the pGEX expression system. Two antibody-binding regions were identified on the basis of the reactivity of a series of truncated gL constructs with anti-MCMV antibodies. One was mapped to residues 1 to 38 and the other between residues 230 and 274. Polyclonal antibodies specific to gL were raised against the full-length gL fusion protein. The antisera were shown to react with a 46K protein present in purified virions by Western blotting. Treatment of purified virions with endoglycosidase-H or -F resulted in reductions in M(r) of the 46K species to 42K and 31K, respectively. The antisera did not exhibit any neutralizing activity in a plaque reduction assay.
对小鼠巨细胞病毒（MCMV）史密斯株基因组的DNA序列分析显示，有一个开放阅读框（ORF），其氨基酸序列与其他疱疹病毒的糖蛋白L（gL）具有同一性。该ORF大小为822 bp，能够编码一个含274个氨基酸的蛋白质。它与人类巨细胞病毒和人类疱疹病毒6的gL基因具有显著同一性。还确定了MCMV K181株gL基因的编码序列，并使用pGEX表达系统在大肠杆菌中作为与谷胱甘肽S-转移酶的融合蛋白进行表达。基于一系列截短的gL构建体与抗MCMV抗体的反应性，鉴定出两个抗体结合区域。一个定位在第1至38位氨基酸残基，另一个在第230至274位氨基酸残基之间。针对全长gL融合蛋白制备了gL特异性多克隆抗体。通过蛋白质免疫印迹法显示，抗血清与纯化病毒粒子中存在的一种46K蛋白发生反应。用内切糖苷酶-H或-F处理纯化的病毒粒子，导致46K蛋白的相对分子质量分别降至42K和31K。在蚀斑减少试验中，抗血清未表现出任何中和活性。
