重庆医学2017，Vol.46Issue(5)：583-585,3.DOI:10.3969/j.issn.1671-8348.2017.05.003
大鼠Hes1腺病毒表达载体构建及功能鉴定
Construction and functional identification of rat Hes1 adenovirus expression vector
摘要
Abstract
Objective to construct the high titers rat Hes1 adenovirus expression vector (Ad-Hes1).Methods With the rat cDNA as a template,the Hes1 fragment was amplified by PCR,which constructed pShuttle-CMV-Hes1 shuttle plasmid by directly clone.Based on pShuttle-CMV-Hes1,pAdeno-Hes1 virus plasmid was constructed,pAdeno-Hes1 was transfected into 293 cells to package Ad-Hes1,virus titers were determined by modified TCID50.Hes1 was detected by Western blot after Ad-Hes1 infected with H9c2 myocardial cells.Results pShuttle-CMV-Hes1 shuttle plasmid and pAdeno-Hes1 plasmid were constructed successfully,with a general titer of 1.6 × 1011 PFU,Ad-Hes1 can be expressed in H9c2 myocardial cells,and its MOI value was 30.Conclusion Ad-Hes1 is successfully constructedand packaged,thus provide basis for further research on the protection effect of Hes1 on myocardium.关键词
腺病毒科/肌细胞,心脏/Hes1/质粒构建/病毒包装Key words
adenoviridae/myocytes, cardiac/Hes1/plasmid construction/virus packaging分类
医药卫生引用本文复制引用
周学亮,方义湖,赵勇,邹斌,徐华,刘季春..大鼠Hes1腺病毒表达载体构建及功能鉴定[J].重庆医学,2017,46(5):583-585,3.基金项目
国家自然科学基金资助项目(81570262) （81570262）
赣鄱555领军人才计划(赣组字[2013]58号) （赣组字[2013]58号）
江西省自然科学基金重大项目(20152ACB20026). （20152ACB20026）
