解放军医学院学报Issue(3)：258-260,3.DOI:10.3969/j.issn.2095-5227.2014.03.018
Gateway技术构建转基因重组质粒pRP.EX3d-EF1A-LRP16-His-IRES-eGFP
Application of gateway technology in construction of recombinant transgenic vector pRP. EX3d-EF1A-LRP16-His-IRES-eGFP
摘要
Abstract
Objective To lay the foundation for establishing transgenic mice by constructing recombinant transgenic vector pRP. EX3d-EF1A-LRP16-His-IRES-eGFP. Methods The attB1- LRP16-His-attB2 was amplified by overlap PCR. The LRP16 gene was cloned into pDown vector via BP reaction and transmitted into pRP.EX3d vector via LR reaction with pDown-LRP16-His, pUp-EF1A, pTail-IRES-eGFP and PRP. Des3d. The recombinant plasmid was transformed into Stbl3 cells and screened using the ampicillin (AMP) resistance gene. Positive clones were confirmed by PCR and DNA sequencing, respectively. Results Sequencing and restriction endonuclease showed that the recombinant vector pRP.EX3d-EF1A-LRP16-His-IRES-eGFP was successfully constructed. Conclusion The successfully constructed recombinant vector pRP.EX3d-EF1A-LRP16-His-IRES-eGFP can be used in establishing LRP16 transgenic mice.关键词
LRP16/转基因小鼠/质粒/Gateway技术Key words
LRP16/transgenic mice/plasmid/gateway method分类
医药卫生引用本文复制引用
柏苗苗,王春萌,伍志强,梅倩,李小雷,李祥,赵亚力,韩为东..Gateway技术构建转基因重组质粒pRP.EX3d-EF1A-LRP16-His-IRES-eGFP[J].解放军医学院学报,2014,(3):258-260,3.基金项目
国家自然科学基金项目(31270820；81230061；81072109)；北京市科技新星计划(2011113)@@@@Supported by the National Natural Science Foundation of China(31270820 （31270820；81230061；81072109）
81230061 （）
81072109) （）
Beijing Nova Program (2011113) （2011113）
