摘要
目的建立瞬时受体电位通道家族M8型受体(TRPM8)转基因小鼠模型。方法通过显微注射法将pcDNA3.1-h Trpm8导入C57BL/6J×CBA F1小鼠的受精卵中,并将其移植到假孕母鼠输卵管中获得子代小鼠。采用PCR方法鉴定子代基因型,通过RT-PCR和免疫印迹法检测Trpm8基因及蛋白在组织中的表达。结果将405枚注射受精卵移植给15只假孕鼠,其中13只顺利妊娠并产下95只子鼠,经PCR鉴定得到5只转基因阳性首建鼠,其中4只小鼠可稳定传代,经与C57BL/6J小鼠回交7代后互交数代建系。子代转基因小鼠经RT-PCR检测发现Trpm8基因在心脏、肝脏、肾脏、脂肪和骨骼肌中均有表达,且TRPM8蛋白表达在转基因小鼠中显著增加。结论通过显微注射法成功建立了Trpm8转基因小鼠模型。
Objective To establish an animal model of transient receptor potential melastatin 8（TRPM8）channel transgenic mice.Methods pcDNA3.1-hTrpm8 was injected into the fertilized mouse eggs from C57BL/6J ×CBA F1 mice.The injected eggs were transplanted into the oviduct of pseudopregnant mice.Tail DNA PCR screening was performed to identify the positive founder mice.The expressions of Trpm8 mRNA and protein in the tissues of the transgenic mice were detected by RT-PCR and Western blotting.Results 405 eggs were transplanted into the oviducts of 15 recipients.95 viable offspring were born from 13 of the recipients.Tail DNA PCR screening showed 5 of the offspring were positive transgenic mice.As detected by RT-PCR,Trpm8 mRNA expressions were present in the heart,liver,kidney,adipose tissue and skeletal muscle of the transgenic mice.Additionally,the TRPM8 protein expression was significantly increased in the transgenic mice.Conclusion TRPM8 transgenic mice have been successfully established.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2010年第4期251-254,共4页
Journal of China Medical University
基金
国家自然科学基金资助项目(30670839)
