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《Chongqing Medicine》 2017-05
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Construction and functional identification of rat Hes1 adenovirus expression vector

Zhou Xueliang;Fang Yihu;Zhao Yong;Zou Bin;Xu Hua;Liu Jichun;Department of Cardiovascular Surgery,the First Affiliated Hospital of Nanchang University;  
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 constructed and packaged,thus provide basis for further research on the protection effect of Hes1 on myocardium.
【Fund】: 国家自然科学基金资助项目(81570262);; 赣鄱555领军人才计划(赣组字[2013]58号);; 江西省自然科学基金重大项目(20152ACB20026)
【CateGory Index】: Q78
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