肝组织特异表达人核受体nr5a2转基因小鼠的构建

人乙肝病毒增强子ⅡB1结合因子（hB1F）系Ftz—F1（NR5A）亚家族的新成员。经基因重组法将人hb1 fcDNA置于小鼠白蛋白增慢子／启动子序列下游构建成肝特异重组载体，通过原核显微注射将该载体导入小鼠受精卵原核，经注射且状态良好的卯回输至假孕母鼠输卯管。产下仔鼠经PCR和Southern blotting鉴定，同时RT—PCR和Western blotting分析转基因的表达。阳性Founder鼠与正常C57鼠交配以建立转基因纯系小鼠，F1代以PCR法鉴定。结果共获得4只PCR鉴定转基因阳性Founder鼠，其中一只同时经Southern blotting鉴定为阳性。RT—PCR和Western blotting结果显示，外源基因在转基因小鼠的肝组织成功表达。遗传学分析表明，转基因已整合入小鼠基因组并可稳定溃传。

Generation of Transgenic Mice with Liver-specific Expression of Human Nuclear Receptor nr 5a 2

Human nuclear receptor hb1f(nr5a2) was cloned and characterized as a novel member of the Ftz-F1(nr5a) nuclear receptor subfamily,whose its biological function remained largely unidentified.The aim of this study was to establish transgenic mouse model that specifically expressed hB1F in the liver to faciliate the functional study of hB1F.hb1f cDNA was placed downstream of mouse albumin gene enhancer/promoter to construct a liver-specific hb1f expression vector.Transgene fragments were microinjected into fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.Four offspring were identified as carrying the transgenes by PCR,from which one was also verified by Southern blotting.RT-PCR and Western blotting results showed that the transgene was expressed in the liver of the transgenic mice.Transgenic founder mice were used to establish transgenic mouse lineages.The F1 mice were identified by PCR analysis.Genetic analysis of the transgenic mice demonstrated that the transgene had been integrated into the chromosome at a single site and could be stably transmitted.