Wang Shui-Liang, Yang Hua, Xie You-Hua, Wang Yuan, Li Jian-Zhong, Wang Long, Wang Zhu-Gang, Fu Ji-Liang
Department of Medical Genetics, Second Military Medical University, Shanghai 200433, China.
Yi Chuan Xue Bao. 2005 Dec;32(12):1241-7.
Human nuclear receptor hb1 f(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.
人类核受体hb1f（nr5a2）被克隆并鉴定为Ftz-F1（nr5a）核受体亚家族的一个新成员，但其生物学功能在很大程度上仍不明确。本研究的目的是建立在肝脏中特异性表达hB1F的转基因小鼠模型，以促进hB1F的功能研究。将hb1f cDNA置于小鼠白蛋白基因增强子/启动子下游，构建肝脏特异性hB1F表达载体。将转基因片段显微注射到小鼠受精卵中。将操作后的胚胎移植到假孕雌性小鼠的输卵管中。通过PCR鉴定出4只携带转基因的后代，其中1只也通过Southern印迹法得到验证。RT-PCR和Western印迹结果表明，转基因在转基因小鼠的肝脏中表达。转基因奠基小鼠用于建立转基因小鼠品系。通过PCR分析鉴定F1小鼠。对转基因小鼠的遗传分析表明，转基因已整合到染色体的单个位点，并且可以稳定遗传。
