中国医科大学学报2017，Vol.46Issue(6)：489-494,500,7.DOI:10.12007/j.issn.0258⁃4646.2017.06.003
人巨细胞病毒Han株US12基因缺失株的构建及其在人胚肺成纤维细胞中的复制
Construction of Human Cytomegalovirus Han?ΔUS12?BAC Strain and Studies on Its Replication in Human Embryonic Lung Fibroblasts
摘要
Abstract
Objective To construct human cytomegalovirus(HCMV)Han?ΔUS12?BAC strain and to study the role of US12 in HCMV replica?tion in human embryonic lung fibroblasts(HELF). Methods Kanamycin?resistant gene was amplified with primers containing homology arms se?quence flanking US12 and then electroporated into E.coli DY380?Han?wt?BAC competent cells. Successfully recombinant Han?ΔUS12?BAC clones were identified by PCR,sequenced for confirmation Han?ΔUS12?BAC plasmids were electroporated into HELF to produce infectious viri?ons. Han?ΔUS12?BAC and Han?wt?BAC were inoculated onto HELF at the multiplicity of infection of 0.1 pfu/cell. The viral titer in the supernatant was measured by TCID50 assay and growth kinetics of the viruses in HELF was studied. Results Han?ΔUS12?BAC clone was successfully con?structed. Han?ΔUS12?BAC was reconstructed in HELF to generate infectious virions. Growth kinetics assay indicated that Han?ΔUS12?BAC and Han?wt?BAC showed no differences in their growth and dissemination in HELF. Conclusion US12 in HCMV clinical strain Han is dispensable for HCMV growth and dissemination in HELF.关键词
巨细胞病毒/US12/细菌人工染色体/生长曲线Key words
cytomegalovirus/US12/bacterial artificial chromosome/growth kinetics分类
医药卫生引用本文复制引用
卢颖,黄郁晶,阮强,马艳萍,柳中洋,王鸿雁,齐莹,韩丽英,高爽,郑博,刘畅..人巨细胞病毒Han株US12基因缺失株的构建及其在人胚肺成纤维细胞中的复制[J].中国医科大学学报,2017,46(6):489-494,500,7.基金项目
国家自然科学基金(30672248,81371788) （30672248,81371788）
盛京医院自由研究者项目 （）
