BAMBI基因敲除小鼠的繁育、基因型鉴定

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.

Breeding and Identification of BAMBI Gene Knockout Mice

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 prog- enies 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.