Pande H, Campo K, Tanamachi B, Zaia J A
Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California 91010.
Virology. 1991 Oct;184(2):762-7. doi: 10.1016/0042-6822(91)90448-k.
Human cytomegalovirus (HCMV) contains a 28-kDa (pp28) matrix phosphoprotein which has been shown to be highly immunogenic in humans. We have cloned and sequenced the gene encoding pp28 of HCMV Towne strain (pp28Towne) and have expressed this gene in stable Chinese hamster ovary (CHO) cell lines in order to examine the structural, functional, and antigenic properties of this protein. The pp28Towne gene had 99% nucleotide and 98.4% amino acid similarity to the pp28 gene of HCMV AD169 strain (pp28AD169). We identified three amino acid substitutions (Gly70 to Ser70, Ser76 to Asn76, and Thr85 to Ala85) in pp28Towne, all clustered in a short 16 amino acid stretch located in the N-terminal half of the protein. The pp28Towne gene was expressed in CHO cells using a vector in which transcription was driven by a human beta-actin promoter. The expressed protein, having an electrophoretic mobility similar to that of HCMV-derived pp28, reacted strongly in immunoblot analysis with pp28-specific murine monoclonal antibodies as well as HCMV-seropositive human sera.
人巨细胞病毒（HCMV）含有一种28 kDa（pp28）的基质磷蛋白，已证明该蛋白在人体内具有高度免疫原性。我们克隆并测序了HCMV汤氏株（pp28Towne）编码pp28的基因，并在稳定的中国仓鼠卵巢（CHO）细胞系中表达了该基因，以研究该蛋白的结构、功能和抗原特性。pp28Towne基因与HCMV AD169株（pp28AD169）的pp28基因具有99%的核苷酸相似性和98.4%的氨基酸相似性。我们在pp28Towne中鉴定出三个氨基酸替换（Gly70变为Ser70、Ser76变为Asn76以及Thr85变为Ala85），所有这些替换都集中在该蛋白N端一半的一段短16个氨基酸区域内。使用由人β-肌动蛋白启动子驱动转录的载体，在CHO细胞中表达pp28Towne基因。表达的蛋白在免疫印迹分析中与pp28特异性鼠单克隆抗体以及HCMV血清阳性人血清发生强烈反应，其电泳迁移率与HCMV来源的pp28相似。
