临床低传代人巨细胞病毒株UL148开放阅读框mRNA的表达与鉴定  

Identification and expression of human cytomegalovirus UL148 mRNA of low-passage clinical isolates in Guangzhou

作　　者：王波[1] 李月琴[2] 叶宁[1] 胡兢晶[1] 苏海浩[1] 何震宇[2] 周天鸿[2] 

机构地区：[1]广东省妇幼保健院儿科,广州510010 [2]暨南大学生命科学与技术学院

出　　处：《中华医学杂志》2008年第24期1693-1696,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30370776）;广东省自然科学基金资助项目（06022095）;广东省医学科学基金资助项目（2006280）;广州市教育局科技计划项目基金资助项目（2004-1045）

摘　　要：目的研究广州地区人巨细胞病毒（HCMV）临床病毒株特有基因UL148的结构，分析其结构多态性与HCMV先天性感染致病的关系；研究UL148的表达情况，探讨其产物可能的功能及在致病机制中的可能作用。方法采集广州地区新生儿HCMV感染患者尿液标本进行病毒分离培养，从分离的病毒株中提取病毒DNA，扩增鉴定UL148基因片段；TA克隆，基因测序；抽提病毒总RNA，用RT-PCR方法鉴定UL148基因mRNA表达情况。结果成功分离到HCMV低传代临床株，获得UL148基因全序列。RT-PCR产物检测582bp大小的特异性条带，证实了UL148基因mRNA的表达。结论广州地区临床低传代HCMV分离病毒株存在UL148基因结构，证实UL148开放阅读框是具有mRNA水平表达的基因结构。Objective To research the structure of the unique gene UL148 of the human cytomegalovirus （HCMV） clinical strain in Guangzhou, analyze the relation between its structure polymorphism and HCMV congenital infection, investigate the function of its expression products, and try to reveal the pathogenic mechanism of the gene in HCMV infection in newborns. Methods Urine samples were collected from 10 newborns with HCMV infection and inoculated on human fetal lung cells. The viral DNA was isolated, and UL148 gene fragment was amplified and identified. After TA clone and gene sequencing, total RNA of virus was extracted and the mRNA expression of UL148 gene was identified by using RT-PCR. Results The UL148 gene complete sequence was determined. A specific 582 bp length DNA fragment was amplified by RT-PCR, which confirmed the expression of UL148 gene. Conclusions UL148 gene exists in the low-passage clinical HCMV isolate from Guangzhou. It is confirmed that open reading frame of UL148 has genetic structure with mRNA expression.

关 键 词：巨细胞病毒 RNA 信使 低传代 

分 类 号：R686[医药卫生—骨科学]