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表达大鼠FoxA1的慢病毒制备和鉴定
引用本文:谭拥军,谢骊,杨潮,何斯佳,谭桂湘. 表达大鼠FoxA1的慢病毒制备和鉴定[J]. 湖南大学学报(自然科学版), 2012, 39(3): 58-61
作者姓名:谭拥军  谢骊  杨潮  何斯佳  谭桂湘
作者单位:湖南大学生物学院
基金项目:国家自然科学基金资助项目(30871244);科技部国际合作重大资助项目(2010DFB30300)
摘    要:通过PCR扩增获得大鼠FoxA1的cDNA,构建慢病毒重组载体plv-rFoxA1-IRES-EGFP,并瞬时转染293T细胞观察绿色荧光蛋白(GFP)的表达及Western blot检测FoxA1的表达.通过钙转将该重组载体与辅助质粒△8.91,pVSV-G共转染293T细胞制备慢病毒.研究结果表明,大鼠FoxA1慢病毒质粒成功构建,并且证实所制备的慢病毒能感染细胞,使细胞有效表达FoxA1蛋白,为FoxA1的功能研究奠定了材料基础.

关 键 词:质粒  慢病毒  FoxA1基因  PCR

Construction and Identification of the Lentivirus Expressing Rat FoxA1
TAN Yong-jun,XIE Li,YANG Chao,HE Si-ji,TAN Gui-xiang. Construction and Identification of the Lentivirus Expressing Rat FoxA1[J]. Journal of Hunan University(Naturnal Science), 2012, 39(3): 58-61
Authors:TAN Yong-jun  XIE Li  YANG Chao  HE Si-ji  TAN Gui-xiang
Affiliation:(School of Biology,Hunan Univ,Changsha,Hunan 410082,China)
Abstract:The coding region of FoxA1,amplified by PCR,was cloned to construct the lentivirus vector plv-rFoxA1-IRES-EGFP.The recombinant plasmid was transfected transiently into 293T cells and the expressions of GFP and FoxA1 were confirmed by fluorescent microscopy and Western blot.The plv-rFoxA1-IRES-EGFP was cotransfected with the packaging plasmids △8.91,pVSV-G to 293T cells to produce the lentivirus.The results show that lentiviral vector that expresses rat FoxA1 was successfully constructed,and infection ability of the recombinant lentivirus was confirmed by the effective expression of FoxA1 protein in cells.These results provided an important material to study the FoxA1 functions in the future.
Keywords:plasmid  lentivirus  FoxA1  PCR
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