BAMBI基因敲除小鼠的繁育、基因型鉴定

doi:10.3969/j.issn.1674-5817.2014.05.007

实验动物与比较医学 ›› 2014, Vol. 34 ›› Issue (5): 381-385.DOI: 10.3969/j.issn.1674-5817.2014.05.007

所属专题：实验动物资源开发与利用

BAMBI基因敲除小鼠的繁育、基因型鉴定

罗肖¹, 魏双羽^1,2, 贾如¹, 姚婷¹, 张燕茹¹, 师小博¹, 闫剑群¹

1.西安交通大学医学部生理学与病理生理学系; 教育部环境与疾病相关基因重点实验室,西安 710061;
2.西安交通大学第一附属医院血液内科, 西安 710061

收稿日期:2014-03-19 出版日期:2014-10-25 发布日期:2014-10-25
作者简介:罗肖(1982-), 女, 博士, 生理学讲师, E-mail:xluo@mail.xjtu.edu.cn
基金资助:
国家自然科学基金项目资助(编号:31200886); 中央高校基本科研业务费专项资金资助

Breeding and Identification of BAMBI Gene Knockout Mice

LUO Xiao¹, WEI Shuang-yu^1,2, JIA Ru¹, YAO Ting¹, ZHANG Yan-ru¹, SHI Xiao-bo¹, YAN Jian-qun¹

1. Department of Physiology and Pathophysiology; Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi’an Jiaotong University School of Medicine, Xi’an 710061, China;
2. Department of Hematology, the First Affiliated Hospital of Medical College, Xi’an Jiaotong University, Xi’an 710061, China;

Received:2014-03-19 Online:2014-10-25 Published:2014-10-25

摘要/Abstract

摘要： 目的探讨BAMBI^-/-小鼠的繁殖和鉴定方法,筛选出足够数量的BAMBI^-/-小鼠,为研究BAMBI基因在肥胖、能量代谢等方面的生理功能提供实验动物。方法将引进的2对BAMBI^-/-小鼠扩繁后,以与背景品系C57BL/6J野生型(WT)回交的方式获得BAMBI+/-小鼠,再将BAMBI^+/-小鼠进行互交大量繁殖。提取幼鼠尾部组织DNA,利用PCR方法鉴定基因型;选取BAMBI^-/-小鼠取肩胛部位棕色脂肪组织(BAT),腹股沟部位皮下白色脂肪组织(sWAT),性腺周围白色脂肪组织(gWAT)和肝脏,利用实时 PCR方法检测BAMBI mRNA的表达量。结果 BAMBI+/-小鼠互交扩繁,短期能获得大量小鼠;C57BL/6J野生型(WT)、BAMBI^+/-和BAMBI^-/-各表型结果基本符合孟德尔遗传规律;BAMBI^-/-小鼠敲除效率理想。结论 BAMBI^+/-小鼠互交是繁育BAMBI^-/-小鼠的较好方法;PCR方法能够准确鉴定BAMBI^-/-小鼠。

关键词: BAMBI, 基因敲除, 基因型鉴定, PCR

Abstract: Objective Breed and identify BAMBI knockout mice for studying the role of BAMBI in obesity and energy metabolism. Methods The introduced BAMBI^-/- mice were paired ,and the progenies were back-crossed to their wild type mice (C57BL/6J background). The offspring were inter-crossed to obtain sufficient number of the knockout mice, the genome DNA was extracted from the tail of the mice for genotyping by PCR. The BAMBI mRNA expression in liver, brown adipose tissue, subcutaneous and gonadal white adipose tissue of BAMBI knockout and wild type mice were detected by real time PCR. Results The inter-crossing results of heterozygous mice with BAMBI gene were basically accordant with Mendelian inheritance laws. The BAMBI mRNA expression was hardly detected in BAMBI knockout mice. Conclusion It is feasible to breed BAMBI knockout mice by inter-crossing of the heterozygote. PCR technique can be used to identify the genotype of BAMBI knockout mice precisely.

Key words: BAMBI, Knockout, Genotyping, PCR, Real time PCR

中图分类号:

Q95-33

罗肖, 魏双羽, 贾如, 姚婷, 张燕茹, 师小博, 闫剑群. BAMBI基因敲除小鼠的繁育、基因型鉴定[J]. 实验动物与比较医学, 2014, 34(5): 381-385.

LUO Xiao, WEI Shuang-yu, JIA Ru, YAO Ting, ZHANG Yan-ru, SHI Xiao-bo, YAN Jian-qun. Breeding and Identification of BAMBI Gene Knockout Mice[J]. Laboratory Animal and Comparative Medicine, 2014, 34(5): 381-385.

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BAMBI基因敲除小鼠的繁育、基因型鉴定

Breeding and Identification of BAMBI Gene Knockout Mice

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