中国医药科学Issue(13)：34-37,127,5.
真核表达载体pEGFP-N1-kin17的构建及其在Zmpste24-/-小鼠胚胎成纤维细胞内的表达
Construction pEGFP-N1-kin17 eukaryotic expression vector and its expression inZmpste24-/- mouse embryonic fibroblasts
摘要
Abstract
Objective To construct the kin17 GFP fusion eukaryotic expression vector and explore the expression and location of kin17 inZmpste24-/-mouse embryonic fibroblasts (MEFs).Methods cDNA fragment of kin17 was amplified from pCMV-kin17 by PCR and cloned into GFP fusion eukaryotic expression vector pEGFP-N1. The positive clone was confirmed by sequencing.Zmpste24-/- and wild type MEFs were obtained from 13.5 d pregnant Zmpste24+/- female mouse. The recombinant plasmid was transiently transfected intoZmpste24-/- MEFs with LipofectamineTM2000. The protein expression of kin17 was detected by fluorescence microscopy.Results Eukaryotic expression vector pEGFP-N1-kin17 containing coding region of human kin17 gene was successfully constructed. Zmpste24-/- and wild type MEFs were identified by PCR and western blotting genotyping.Zmpste24-/- and wild type MEFs transfected with the recombinant plasmid expressed high level of recombinant kin17 protein was detected by fluorescence microscopy.ConclusionThe construction of pEGFP-N1-kin17 was successfully achieved and the expression of GFP-kin17 was located in the nucleus ofZmpste24-/- and wild type MEFs.关键词
kin17/GFP/Zmpste24/小鼠胚胎成纤维细胞Key words
kin17/GFP/Zmpste24/Mouse embryonic fibroblasts分类
医药卫生引用本文复制引用
郑慧玲,崔红晶,余磊,黄颖,刘新光..真核表达载体pEGFP-N1-kin17的构建及其在Zmpste24-/-小鼠胚胎成纤维细胞内的表达[J].中国医药科学,2014,(13):34-37,127,5.基金项目
国家自然科学基金资助项目（30672205，81170327）；广东省自然科学基金重点项目（9252402301000002）。 （）
