文章摘要
周建丽,赖娅娜,李树珍,曾文滔.pCAG⁃3×flag⁃PP2A Cα转基因小鼠构建及高表达系的筛选[J].南京医科大学学报,2018,(3):283~287
pCAG⁃3×flag⁃PP2A Cα转基因小鼠构建及高表达系的筛选
Constructing of pCAG⁃3×flag⁃PP2A Cα transgenic mice and screening of high level expression system
投稿时间:2017-03-30  
DOI:10.7655/NYDXBNS20180301
中文关键词: pCAG⁃3×flag⁃PP2A Cα  转基因小鼠  过表达
英文关键词: pCAG⁃3×flag⁃PP2A Cα  transgenic mice  over⁃expression
基金项目:国家自然科学基金(31171443)
作者单位
周建丽 南京医科大学医药实验动物中心医药模式动物重点实验室江苏 南京 211166 
赖娅娜 南京医科大学医药实验动物中心医药模式动物重点实验室江苏 南京 211166 
李树珍 南京医科大学医药实验动物中心医药模式动物重点实验室江苏 南京 211166 
曾文滔 南京医科大学医药实验动物中心医药模式动物重点实验室江苏 南京 211166 
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中文摘要:
      目的:构建过表达pCAG?3×flag?PP2A Cα转基因小鼠,筛选高表达稳定遗传系,建立PP2A Cα基因相关功能研究模型动物。方法:将Flag蛋白标签序列与鼠源PP2A Cα cDNA 转录本设计成融合基因,插入CAG启动子下游,构建过表达载体pCAG?3×flag?PP2A Cα。通过原核显微注射技术,将线性化pCAG?3×flag?PP2A Cα质粒注射入受精卵雄原核,构建pCAG?3×flag?PP2A Cα过表达模型小鼠,PCR筛选阳性过表达小鼠。提取阳性过表达小鼠组织总蛋白,在蛋白质水平分析转基因小鼠3×flag?PP2A Cα的表达。结果:PCR 鉴定及测序结果证实,pCAG?3×flag?PP2A Cα 转基因载体及过表达pCAG?3×flag?PP2A Cα 转基因小鼠模型构建成功。Western blot筛选转基因高表达系小鼠。结论:筛选得到pCAG?3×flag?PP2A Cα高表达稳定遗传系小鼠。
英文摘要:
      Objective:To construct over?expressing pCAG?3×flag?PP2A Cα transgenic mice,screen the stable system of high level expression,and therefore to provide a transgenic animal model for the study of the biological function of PP2A Cα. Methods:The pCAG?3×flag?PP2A Cα transgenic vectors were constructed by inserting mice PP2A Cα cDNA and Flag protein sequence tag to the downstream of pCAG promoter. The linear pCAG?3×flag?PP2A Cα plasmid was injected into the zygote nucleus by microinjection technique to get the over?expressing pCAG?3×flag?PP2A Cα mice. The positive over?expression mice were identified by PCR. The positive expression of total protein in mice was extracted and the expression of transgenic mice was analyzed at the protein level. Results:The pCAG?3×flag?PP2A Cα transgenic vector and transgenic mice was successfully constructed by PCR and was verified by plasmid sequence analysis. The system of high level expression transgenic mice was screened by Western blot assay. Conclusion:The pCAG?3×flag?PP2A Cα transgenic mice with stable system of high level expression were screened and obtained.
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