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RANTES-K慢病毒载体的构建及其慢病毒的产生和鉴定
引用本文:孙利,张颖,黄长形,张久聪,连建奇,张野,魏欣,孙永涛,白雪帆.RANTES-K慢病毒载体的构建及其慢病毒的产生和鉴定[J].第四军医大学学报,2009,30(3):247-250.
作者姓名:孙利  张颖  黄长形  张久聪  连建奇  张野  魏欣  孙永涛  白雪帆
作者单位:第四军医大学唐都医院全军感染病诊疗中心,陕西,西安,710038  
摘    要:目的:构建含CC细胞内趋化因子(CC-intrakine,RANTES-K)基因的慢病毒载体,为研究Ⅰ型人免疫缺陷病毒(HIV-1)感染的基因治疗奠定基础.方法:应用PCR技术,扩增目的基因片段RANTES-K,纯化后的PCR产物定向连接入pLenti6/V5-D-TOPO载体,构建慢病毒载体pLenti6/V5-R-K,并在293FT细胞中建立慢病毒株,最后转染HeLa细胞观察RANTES蛋白的表达.结果:成功构建了慢病毒载体pLenti6/V5-R-K,并证实RANTES蛋白可在人宫颈癌HeLa细胞系内表达.结论:慢病毒载体可介导CC-intrakine(RANTES-K)基因高效稳定转染HeLa细胞,可用于抗HIV-1感染的基因治疗.

关 键 词:HIV-1  趋化因子受体  细胞内趋化因子  慢病毒载体  基因治疗

Construction and expression of pLenti6/V5-R-K with CC-intrakine
SUN Li,ZHANG Ying,HUANG Chang-Xing,ZHANG Jiu-Cong,LIAN Jian-Qi,ZHANG Ye,WEI Xin,SUN Yong-Tao,BAI Xue-Fan PLA Center of Diagnosis , Treatment for Infectious Diseases,Tangdu Hospital,Fourth Military Medical University,Xi''an ,China.Construction and expression of pLenti6/V5-R-K with CC-intrakine[J].Journal of the Fourth Military Medical University,2009,30(3):247-250.
Authors:SUN Li  ZHANG Ying  HUANG Chang-Xing  ZHANG Jiu-Cong  LIAN Jian-Qi  ZHANG Ye  WEI Xin  SUN Yong-Tao  BAI Xue-Fan PLA Center of Diagnosis  Treatment for Infectious Diseases  Tangdu Hospital  Fourth Military Medical University  Xi'an  China
Institution:SUN Li,ZHANG Ying,HUANG Chang-Xing,ZHANG Jiu-Cong,LIAN Jian-Qi,ZHANG Ye,WEI Xin,SUN Yong-Tao,BAI Xue-Fan PLA Center of Diagnosis and Treatment for Infectious Diseases,Tangdu Hospital,Fourth Military Medical University,Xi'an 710038,China
Abstract:AIM: To study the HIV-based lentiviral vector,pLenti6/V5-R-K,which contained the gene CC-intrakine(RANTES-K) and to provide a basis for the gene therapy of HIV-1 infection.METHODS:A pLenti-based expression vector,pLenti6/V5-D-TOPO,was used to produce the lentiviral vector,which was cotransfected with the ViraPowerTM Packaging Mix(pLP1,pLP2,and pLP/VSVG) into 293FT cells to produce a replication-incompetent lentivius stock.After titrating the lentiviral stock using HeLa cells,the expression of the interest g...
Keywords:HIV-1
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