CAJ | 학술논문

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.
목적 구건표체인류소분자열휴극단백22(HSP22)적전기인소서모형. 방법 구건휴대인류HSP22기인적pCAGGS-H A-Wt HSP22전기인표체재체,용Sal Ⅰ、HindⅢ화BsaⅪ3개핵산내절매대pCAGGS-HA-wt HSP22진행매절득도선성화목적편단,통과미섬유주사기술진행수정란원핵주사,통과PCR결합측서기술대자서진행감정,응용Western Blot기술진행외원성HSP22단백표체적분석. 결과 획득4지휴대인류HSP22기인적전기인수건서,분별위Tg646、Tg648、Tg649화Tg661계수건서,Tg661계수건서불표체인류HSP22단백,Tg646、Tg648、Tg649계수건서표체인류HPP22단백,가용우하일보연구. 결론 획득료표체인류HSP22단백적전기인소서모형,위개전HSP22기인공능연구전정료기출.
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.