LSS基因敲除小鼠的饲养、繁殖及基因型鉴定  

Breeding,reproduction and genotype identification of LSS knockout mice

作　　者：李名聪 孙晓梅 李昱昊 周宏 罗欣[1] 张胜权[1] Li Mingcong;Sun Xiaomei;Li Yuhao(Dept of Biochemistry and Molecular, Anhui Medical University, Hefei 230032)

机构地区：[1]安徽医科大学生物化学与分子生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2021年第8期1335-1338,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271748);安徽省高等学校自然科学研究项目(编号:KJ2017A195)。

摘　　要：探讨羊毛固醇合成酶(LSS)基因敲除鼠的繁育和基因型分析。将引进杂合子小鼠进行饲养并繁育,提取其基因组DNA,利用PCR反应扩增目的基因片段,鉴定出小鼠的基因型;Western blot分析并比较LSS基因敲除杂合子小鼠和野生型小鼠的肝脏、皮肤组织中LSS蛋白的表达情况。成功的繁育出LSS基因敲除小鼠,使用PCR鉴定其子代小鼠包括野生型(LSS^(+/+),Wt)、杂合子(LSS^(+/-)),未出现纯合敲除小鼠;Western blot结果表明:相较于野生型小鼠,LSS基因敲除杂合子小鼠在肝组织和皮肤组织中表达LSS蛋白量少;杂合子敲除鼠的生存能力比野生型小鼠弱。获得LSS基因敲除小鼠,PCR鉴定小鼠基因型具有简单、稳定的优点。To explore the breeding and genotype analysis of Lanosterol synthase(LSS)gene knockout mice.The introduced heterozygous knockout mice were reared and bred,the tails of progeny mice around 2 weeks old were cut,their genomic DNA was extracted,the target gene fragments were amplified by PCR reaction and identified by agarose gel electrophoresis Genotype of mice;Western blot analysis was used to analyze and compare LSS protein expression in livers and skin tissues of heterozygous and wild-type mice.A batch of LSS knockout mice were successfully bred,and their progeny mice were identified by PCR with two genotypes:wild type(LSS^(+/+),Wt)and heterozygous(LSS^(+/-)),which did not appear Homozygous knockout mice;Western blot results showed that compared with wild-type mice,LSS gene knockout heterozygous mice expressed less LSS protein in liver and skin tissues;the viability of heterozygous knockout mice was weaker than that of wild-type mice.Successfully obtaining LSS gene knockout mice,PCR identification of mouse genotypes has the advantages of fast,simple and stable,which lays the foundation for further study of the biological function of LSS.

关 键 词：LSS 基因敲除小鼠 基因型鉴定 聚合酶链式反应体系 繁育 

分 类 号：R332[医药卫生—人体生理学]