表达人类小分子热休克蛋白22转基因小鼠模型的建立
Establishment of transgenic mouse models expressing human HSP22 protein
目的 构建表达人类小分子热休克蛋白22(HSP22)的转基因小鼠模型. 方法 构建携带人类HSP22基因的pCAGGS-H A-Wt HSP22转基因表达载体,用Sal Ⅰ、HindⅢ和BsaⅪ3个核酸内切酶对pCAGGS-HA-wt HSP22进行酶切得到线性化目的片段,通过微纤维注射技术进行受精卵原核注射,通过PCR结合测序技术对仔鼠进行鉴定,应用Western Blot技术进行外源性HSP22蛋白表达的分析. 结果 获得4只携带人类HSP22基因的转基因首建鼠,分别为Tg646、Tg648、Tg649和Tg661系首建鼠,Tg661系首建鼠不表达人类HSP22蛋白,Tg646、Tg648、Tg649系首建鼠表达人类HPP22蛋白,可用于下一步研究. 结论 获得了表达人类HSP22蛋白的转基因小鼠模型,为开展HSP22基因功能研究奠定了基础.
更多Objective To establish transgenic mouse models expressing human HSP22 protein.Methods pCAGGS-HA-Wt HSP22 transgenic expressing vector carrying human HSP22 gene was constructed by gene recombination technology.The linearized DNA was got by SalI、Hind Ⅲ and BsaⅪ digestion of PCAGGS-HA-Wt HSP22,purified and microinjected into fertilized eggs from C57BL mice.The tail DNA of pups was tested by PCR and DNA sequencing.Expression of human HSP22 protein was detected by western blot with anti-HA tag monoclonal antibody.Results 4 transgenic founder mice (Tg646,Tg648,Tg649,Tg661) carrying human HSP22 gene were identified by PCR and DNA sequencing.The human HSP22 protein was expressed in the lines Tg646,Tg648 and Tg649 founder mice,but was not expressed in the line Tg661 founder mouse.Conclusions The mouse models expressing human HSP22 protein are established successfully and provide the foundation for HSP22 gene research in vivo.
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