Qian G, Wood C, Chandran B
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City 66160.
Virology. 1993 May;194(1):380-6. doi: 10.1006/viro.1993.1272.
Sequence analysis of an 8-kb BamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephs et al., J. Virol. 65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In the in vitro transcription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. These in vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from [35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.
对来自人类疱疹病毒6型A变种GS株（HHV - 6A(GS)）的一个8kb BamHI - G DNA片段进行的序列分析，鉴定出一个编码与其他疱疹病毒糖蛋白gH同源的糖蛋白的基因，该基因被命名为HHV - 6 gH（约瑟夫斯等人，《病毒学杂志》65卷，5597页）。通过聚合酶链反应扩增了HHV - 6A(GS) gH基因的开放阅读框（ORF），并在细菌中作为trpE融合蛋白表达。仅用包含ORF中第15至469位氨基酸的构建体产生了稳定的融合蛋白，该融合蛋白用于制备兔抗血清。在gH基因的体外转录 - 翻译分析中，添加微粒体膜导致未加工的前体加工成约108K的多肽产物。这些体外合成的前体和产物形式被针对gH融合蛋白的兔抗体免疫沉淀。兔抗gH抗体中和了HHV - 6 A变种GS株和B变种Z - 29株的感染性，免疫沉淀反应鉴定出一种病毒体包膜相关的102K多肽为HHV - 6 A和B变种的糖蛋白gH。除了102K糖蛋白外，具有不同部分肽图谱的约58K和164K的非糖基化多肽也始终从[35S]甲硫氨酸标记的HHV - 6A(GS)感染细胞中共同免疫沉淀，但未从HHV - 6B(Z - 29)感染细胞中共同免疫沉淀。HHV - 6两个变种组的两株病毒中糖蛋白gH分子量的相似性以及针对HHV - 6A(GS) gH的兔抗体对两组病毒感染性的中和作用表明，HHV - 6两个变种组中糖蛋白gH具有保守功能。
