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

作　　者：张付峰[1] 卢晓琴[2] 周亚芳[1] 沈璐[3] 江泓[3] 严新翔[3] 唐北沙[3] 

机构地区：[1]中南大学湘雅医院老年科,长沙410008 [2]中南大学湘雅医院急诊科,长沙410008 [3]中南大学湘雅医院神经内科,410008

出　　处：《中华老年医学杂志》2013年第3期333-337,共5页Chinese Journal of Geriatrics

基　　金：国家自然科学基金，教育部博士点新教师青年基金，中南大学自由探索项目

摘　　要：【摘要】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-HA-…HSP22转基因表达载体，用SalI、HindIll和BsaXI3个核酸内切酶对pCAGGS-HA一”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WesternBlot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。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 Bsa XI 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.

关 键 词：热休克蛋白质类 小分子 小鼠 转基因 

分 类 号：R-332[医药卫生]