| 表达FLP重组酶pCEP4-FLP载体的构建及其在FLP/FRT重组系统中的应用 |
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| 引用本文： | 肖雪宾,袁婷,纪元,宋奇,韩晓蕾,武蓉,林晓晨,欧阳红生,李莉.表达FLP重组酶pCEP4-FLP载体的构建及其在FLP/FRT重组系统中的应用[J].河北师范大学学报(自然科学版),2012(3):291-296. |
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| 作者姓名： | 肖雪宾 袁婷 纪元 宋奇 韩晓蕾 武蓉 林晓晨 欧阳红生 李莉 |
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| 作者单位： | 吉林大学 畜牧兽医学院 动物胚胎工程吉林省重点实验室 |
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| 基金项目： | 国家973项目(2011cba01003) |
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| 摘 要： | 构建重组载体pCEP4-FLP和pEF1a-FRT-stop-FRT-GFP,重组载体pEF1a-FRT-stop-FRT-GFP瞬时转染PK-15细胞24h后,再将重组载体pCEP4-FLP瞬时转染该细胞,利用荧光显微镜观察PK-15细胞GFP蛋白的表达情况,利用细胞流式技术统计绿色荧光蛋白表达效率.结果表明:重组载体pCEP4-FLP在细胞内能够表达FLP重组酶,并且表达的FLP重组酶能够识别FRT位点,删除两同向FRT位点间的DNA片段,为FLP/FRT位点特异性重组系统在转基因猪的应用奠定了基础.
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| 关 键 词： | FLP/FRT系统 位点特异性重组 PK-15细胞系 |
Construction of FLP Recombinase Expression Vector pCEP4-FLP and Its Application in FLP/FRT System |
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| XIAO Xuebin,YUAN Ting,JI Yuan,SONG Qi,HAN Xiaolei,WU Rong,LIN Xiaochen,OUYANG Hongsheng,LI Li.Construction of FLP Recombinase Expression Vector pCEP4-FLP and Its Application in FLP/FRT System[J].Journal of Hebei Normal University,2012(3):291-296. |
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| Authors: | XIAO Xuebin YUAN Ting JI Yuan SONG Qi HAN Xiaolei WU Rong LIN Xiaochen OUYANG Hongsheng LI Li |
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| Institution: | (Jilin Provincial Key Laboratory of Animal Embryo Engineering,Institute ofAnimal and Veterinary Medicine,Jilin University,Jilin Changchun 130062,China) |
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| Abstract: | The recombinant vectors pCEP4-FLP and pEF1a-FRT-stop-FRT-GFP were transfected by pCEP4-FLP.The expression of EGFP were analyzed by fluorescence microscope.FACS analysis of the induction of EGFP expression.The recombinant vector pCEP4-FLP can express FLP recombinase which can recognize FRT sites and delete the DNA fra-1gment between two FRT sites in the same orientation.This system will be of great value for the applications of transgenic pigs. |
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| Keywords: | FLP/FRT system site-specific recombinase PK-15 cell line |
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