IP3R2基因敲除小鼠的繁育及基因型鉴定  

Breeding and genotype identification of IP3R2 knockout mice

作　　者：岳雅蓉 项文平 薛慧[3] 赵吉利 王云霞 郐苗 杨琦 YUE Ya-Rong;XIANG Wen-Ping;XUE Hui(Baotou Medical College,Baotou 014040,Inner China)

机构地区：[1]包头医学院,内蒙古包头014040 [2]包头市蒙医中医医院 [3]包头市中心医院 [4]内蒙古医科大学

出　　处：《吉林医学》2024年第10期2319-2323,共5页Jilin Medical Journal

基　　金：国家自然科学基金项目[项目编号:82260451];中央引导地方科技发展资金项目[项目编号:2021ZY0038]。

摘　　要：目的:繁殖并鉴定IP3R2基因敲除小鼠,为后续实验提供样本支持,同时验证PCR法对基因鉴定的适用性。方法:购买IP3R2杂合子小鼠15只,以1雄2雌的合笼方式进行繁殖。繁殖出的子代小鼠生长至6周时,剪子代鼠尾做基因提取及PCR扩增,琼脂糖凝胶电泳结果判定。取鉴定结果为IP3R2基因敲除纯合子小鼠和野生型小鼠脑组织进行Western印迹及免疫荧光双标法,验证脑组织中IP3R2蛋白的表达情况。结果:可成功繁育IP3R2基因敲除小鼠,子代小鼠中,IP3R2纯合率为23.3%。PCR扩增可成功鉴定子代小鼠的基因型。与野生小鼠比较,IP3R2基因敲除小鼠脑组织中IP3R2表达量显著降低,差异有统计学意义(P<0.05)。结论:该实验可成功繁殖并鉴定出IP3R2基因敲除小鼠,为后续实验提供大量样本,PCR技术可快速、准确地鉴定IP3R2基因型,为后续实验提供技术支持。Objective Breeding and identify IP3R2 knockout mice,to provide sample support for the subsequent experiments,and simultaneous validation of PCR applicability for gene identification.Method Fifteen IP3R2 knockout heterozygous mice were purchased and bred in a combined cage of 1 male and 2 females,When the propagated offspring mice were grown to 6 weeks,the shear mouse tail was used for gene extraction and PCR amplification,and the results of agarose gel electrophoresis were determined.Brain samples from homozygous IP3R2 knockout mice and wild-type were indentified for western blot and immunofluorescence to verify the expression of IP3R2 protein in the brain tissue.Daughter mouse brain tissues were harvested for Western blot and immunofluorescence to verify the expression of IP3R2 protein expression in homozygous IP3R2 knockout mice.Results The IP3R2 gene knockout mice can be successfully bred.In the offspring mice,IP3R2 homozygosity was 23.3%.PCR amplification can successfully identify the genotypes of the progeny mice.Compared to the wild mice,IP3R2 expression was significantly lower in IP3R2 knockout mice(P<0.05).Conclusion This experiment successfully established and identified the IP3R2 gene knockout mice,to provide a large number of samples for subsequent experiments,and the PCR technology can quickly and accurately identify the IP3R2 genotype,to provide technical support for the subsequent experiments.

关 键 词：IP3R2 ITPR2 PCR 基因敲除 鉴定 

分 类 号：R3[医药卫生—基础医学]