Lck-Cre×Perp^(flox/flox)条件敲除小鼠的繁育及基因型鉴定  

Reproduction and Genotyping of the Lck-Cre×Perp^(flox/flox) Conditional Knock-Out Mice

作　　者：冷潇 周艳[2] 唐海明[1] 张祥[1] 刘佳[1] 杨佩[1] 王衍堂[1] 

机构地区：[1]成都医学院基础医学院免疫教研室,成都610083 [2]四川大学华西第二医院急诊医学科,成都610041

出　　处：《成都医学院学报》2014年第2期114-117,共4页Journal of Chengdu Medical College

基　　金：国家自然基金资助项目(NO:81202363);四川省教育厅资助项目(NO:13ZB0224);四川省卫生厅资助项目(NO:120257);成都医学院校基金项目(NO:13Z086);成都医学院实验室开放项目(NO:Kf201320)

摘　　要：目的制备Cre-LoxP重组酶系统调控下的T淋巴细胞Perp基因条件性敲除的纯合子小鼠并进行鉴定,为进一步在整体动物水平研究Perp基因在T细胞发育以及在自身免疫疾病发病中的作用提供研究平台。方法将引进的Perp条件敲除小鼠和T淋巴细胞特异性表达Cre重组酶的Lck-Cre小鼠分别进行繁殖和鉴定,然后将两种小鼠进行杂交并对其子代小鼠的基因型进行鉴定,其子代基因型为Lck-Cre×Perpflox/flox的小鼠即为本实验所构建的T细胞Perp基因特异性敲除的纯合子小鼠。结果 Lck-Cre×Perpflox/flox的纯合子小鼠被繁殖成功并准确鉴定。Lck-Cre×Perpflox/flox小鼠脾脏CD3+细胞的Perp基因的RNA及蛋白表达水平与Lck-Cre×Perp+/+小鼠相比显著降低。结论本研究利用Cre-LoxP技术成功构建了T细胞特异性敲除Perp基因的纯合子小鼠,为进一步研究Perp基因在T淋巴细胞发育以及自身免疫性疾病发病中的作用提供了优良的工具。Objective To generate the T lymphocyte-specific Perp knock-out heterozygous mice, for the follow-up research of the role of Perp gene in T cells development and autoimmune disease. Methods The imported Perp conditional knock-out mice and Lck-Cre mice had reproduced and genotyped respectively, then the two strains of mice were crossed, the off-spring mice which genotyping Lck-Cre X Perpflox/flox were the heterozygous mice. Results Lck-Cre 7（ Perpflox/flox mice were breed successfully and their genotype were identified accurately by PCR and western blot. The expression level of Perp in spleen CD3＋ T lymphocytes of Lck-Cre X Perpflox/flox mice was significantly lower, compared with that in Lck-Cre X Perpflox/flox. Conclusion We successfully generated T lymphocyte-specific Perp knock-out heterozygous mice through the Cre-LoxP system, that could be a good tool to study the functions of Perp in T lymphocytes development and autoimmune disease.

关 键 词：Perp Lck-Cre Cre-LoxP系 

分 类 号：R392.5[医药卫生—免疫学]