IDENTIFICATION OF THE MAJOR CAPSID PROTEIN GENE OF HUMAN CYTOMEGALO-VIRUS
IDENTIFICATION OF THE MAJOR CAPSID PROTEIN GENE OF HUMAN CYTOMEGALO-VIRUS
复制标题
DOI：
10.1128/jvi.63.3.1345-1353.1989
复制
发表时间：
1989-03-01
影响因子：
5.4
通讯作者：
JAHN, G
中科院分区：
文献类型：
--
作者：
CHEE, M;RUDOLPH, SA;JAHN, G
将人巨细胞病毒（HCMV）主要衣壳蛋白（MCP）的序列与单纯疱疹病毒（HSV）、eb病毒和水痘带状疱疹病毒的序列进行比较，确定了MCP的编码区。预测的HCMV MCP长度为1370个氨基酸。不同人疱疹病毒的MCP序列与HSV 1型MCP的同源性为25%，与eb病毒MCP的同源性为29%，与水痘-带状疱疹病毒MCP的同源性为23%。编码MCP VP5的HSV 1型KpnI i片段的一个亚片段与含有部分MCP基因的HCMV HindIII U片段交叉杂交。HCMV MCP编码区外亚克隆的Northern （RNA）印迹分析检测到一个约8千碱基的大转录本。一部分开放阅读框在大肠杆菌质粒pBD2 IC2OH中表达为β。-半乳糖苷酶融合蛋白，用于制备新西兰白兔多克隆抗体。获得的抗血清与纯化的HCMV病毒粒子的MCP在Western免疫印迹中反应。人MCP单克隆抗体和兔猴巨细胞病毒Colburn株单特异性抗血清在免疫印迹中检测融合蛋白和纯化病毒粒子的MCP。
The coding region for the major capsid protein (MCP) of human cytomegalovirus (HCMV) was identified by comparing the protein sequence with the respective sequences of herpes simplex virus (HSV), Epstein-Barr virus, and varicella-zoster virus. The predicted length of the HCMV MCP was 1,370 amino acids. Comparison of the MCP sequences of the different human herpesviruses showed a homology of 25% to the MCP of HSV type 1, a homology of 29% to the MCP of Epstein-Barr virus, and a homology of 23% to the MCP of varicella-zoster virus. A subfragment of the HSV type 1 KpnI i fragment encoding the MCP VP5 cross-hybridized with the HCMV HindIII U fragment containing part of the MCP gene. Northern (RNA) blot analyses with subclones out of the coding region for the HCMV MCP detected one large transcript of about 8 kilobases. A portion of the open reading frame was expressed in Escherichia col plasmid pBD2 IC2OH as a .beta.-galactosidase fusion protein and was used to generate polyclonal antibodies in New Zealand White rabbits. The obtained antisera reacted in Western immunoblots with the MCP of purified HCMV virions. A monoclonal antibody against the human MCP and a monospecific rabbit antiserum against strain Colburn of simian cytomegalovirus detected the fusion protein as well as the MCP of purified virions in immunoblots.