类固醇受体辅活化子-3基因敲除小鼠的繁殖与鉴定  

Breeding and identification of SRC-3 knock-out mice

作　　者：李军[1] 粟永萍[1] 王军平[1] 艾国平[1] 刘合年[2] 

机构地区：[1]第三军医大学预防医学院复合伤研究所创伤烧伤与复合伤国家重点实验室,重庆400038 [2]成都军区总医院麻醉科,四川成都610083

出　　处：《西南国防医药》2009年第7期667-669,共3页Medical Journal of National Defending Forces in Southwest China

基　　金：国家重点基础研究发展"973"规划项目(G1999054201);海外青年学者合作研究基金(30328025)

摘　　要：目的:繁殖并鉴定类固醇受体辅活化子(steroid receptor coactivator,SRC)-3基因敲除(SRC-3-/-)小鼠。方法:通过SRC-3-/-雄性小鼠与杂合子(SRC-3+/-)雌鼠杂交,繁殖出SRC-3+/-及SRC-3-/-两种基因型,提取每只小鼠尾部基因组DNA,用PCR法进行鉴定,并进一步提取肝、脾组织总蛋白,采用Western blot进行蛋白表型鉴定。结果:成功繁殖并准确鉴定出更多数目的SRC-3-/-小鼠。SRC-3+/+小鼠脾组织SRC-3蛋白表达水平显著高于肝组织,而SRC-3-/-小鼠的肝、脾组织均未见SRC-3蛋白表达。结论:雄性基因敲除小鼠与雌性杂合子杂交是获取大量SRC-3-/-小鼠的有效途径,PCR法是其子代简单准确的鉴定方法。Objectlve：To breed and identify steroid receptor coaetivator - 3 （SRC -3） knock -out （SRC -3 -/- ） mice. Methods：Male SRC - 3 -/- mice were selected to hybridize with female heterozygote （SRC - 3 ＋/- ） mice to acquire SRC - 3 ＋/- and SRC - 3 -/- baby mice. Genome DNA extracted from the murine tails was subjected to PCR for genotype identification. The total proteins of hepatic and spleen tissue were extracted to identify protein phenotype by Western blot. Results：More SRC - 3 -/- mice were breed successfully by hybridizing male SRC - 3 -/- mice with female heterozygnte （ SRC - 3 ＋/- ） mice and their genotype were identified accurately by PCR. The expression level of SRC - 3 in spleen tissue was significantly higher than that in hepatic tissue in SRC - 3 ＋/＋ mice,but no expression of SRC - 3 was observed in both hepatic and spleen tissue in SRC - 3-/- mice. Conclusion： Hybridization of male gene knock - out mice and female heterozygote mice is an effective way to acquire considerable SRC - 3 -/- mice ,and PCR is a simple and accurate method of identification of genotype.

关 键 词：类固醇受体辅活化子-3 基因敲除 杂交 

分 类 号：Q548.1[生物学—生物化学] R392.13[医药卫生—免疫学]