人类核受体nr5a2在肝脏特异性表达的转基因小鼠的产生。

Shui-Liang Wang, Hua Yang, You-Hua Xie, Yuan Wang, Jian-Zhong Li, Long Wang, Zhu-Gang Wang, Ji-Liang Fu
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引用次数: 0

摘要

人类核受体hb1f (nr5a2)被克隆并鉴定为Ftz-F1 (nr5a)核受体亚家族的新成员,其生物学功能在很大程度上尚未确定。本研究旨在建立肝脏中特异性表达hB1F的转基因小鼠模型,为hB1F的功能研究提供依据。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。将转基因片段微注射到小鼠受精卵中。将经过处理的胚胎移植到假妊娠雌性小鼠的输卵管中。通过PCR鉴定出4个子代携带转基因,其中一个子代也通过Southern blotting进行了验证。RT-PCR和Western blotting结果显示,该基因在转基因小鼠肝脏中有表达。利用转基因创始小鼠建立转基因小鼠系。采用PCR方法对F1小鼠进行鉴定。转基因小鼠的遗传分析表明,该转基因基因已在单个位点整合到染色体中,并能稳定地传播。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Generation of transgenic mice with liver-specific expression of human nuclear receptor nr5a2.

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.

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