中国感染控制杂志2011，Vol.10Issue(2)：81-85,5.
HSV-2多功能蛋白ICP27真核表达载体构建及其在Vero细胞中的表达
Construction of HSV-2 eukaryotic expression plasmid pCDNA3.0-ICP27 and expression in Vero cells
摘要
Abstract
Objective To construct herpes simplex virus type 2(HSV-2) eukaryotic expression plasmid pCDNA3.0-ICP27 and to evaluate its expression in Vero cells. Methods The target sequence of ICP27 gene was obtained and high-fidelity amplified from HSV-2 DNA. The ICP27 gene was cloned into a eukaryote plasmid pCDNA3. 0 after restrictive endonucleases digestion. The pCDNA3. 0-ICP27 was checked and verified by double digestion and DNA sequence analysis. Vero cells were transiently transfected with pCDNA3. 0-ICP27 by lipofectamine 2 000 in vitro.RT-PCR and Western blot analysis were employed to detect the expression of ICP27. Results The 1741bp DNA fragment was obtained by DNA and cloned into pCDNA3. 0. The recombinant plasmid pCDNA3. 0-ICP27 was subjected to sequence analysis which indicated all nucleotides were identical to the HSV-2 ICP27 sequence provided by Genbank. Being transfected by lipofectamine 2 000, the expression of ICP27 in Vero cells was detected. Conclusion Recombinant plasmid pCDNA 3. 0-ICP27 was constructed and expressed successfully in Vero cells.关键词
单纯疱疹病毒2型/疱疹,单纯/感染细胞多肽27/真核表达载体/质粒/疱疹,生殖器分类
医药卫生引用本文复制引用
龙朝钦,杨慧兰,张发洲,薛礼长..HSV-2多功能蛋白ICP27真核表达载体构建及其在Vero细胞中的表达[J].中国感染控制杂志,2011,10(2):81-85,5.基金项目
国家自然科学基金资助项目(30972666)全军医学科学技术研究"十一五"计划资助项目(06J008) （30972666）
