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Establishment of transgenic mice carrying the gene of human nuclear receptor NR5A2 (hB1F)

机译:携带人类核受体NR5A2(hB1F)基因的转基因小鼠的建立

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AIM: Human hepatitis B virus enhancer Ⅱ B1 binding factor (hB1F) was cloned and characterized as a novel member of the Ftz-F1 (NR5A) nuclear receptor subfamily. Although progresses have recently been made, its biological function remains largely unidentified. The aim of this study was to establish an hBlF transgenic mouse model to promote the functional study of hB1F. METHODS: Transgene fragments were microinjected into fertilized eggs of mice. The manipulated embryos were transferred into the oviducts of pseudopregnant female mice. The offsprings were identified by PCR and Southern blot analysis. Transgene expression was analyzed with RT-PCR and Western blot analysis. Transgenic founder mice were used to establish transgenic mouse lineages. The F1 and F2 mice were identified by PCR analysis. RESULTS: Seven mice were identified as carrying copies of transgene. RT-PCR and Western blotting results showed that the transgene was expressed in heart, liver, lung, kidney and stomach in one of the transgenic mouse lineages. Genetic analysis of the transgenic mice demonstrated that the transgene was integrated into the chromosome at a single site, and was transmitted stably. CONCLUSION: In this study we established an hB1F transgenic mouse model, which will facilitate the investigation of the biological function of hB1F in vivo.
机译:目的:克隆人乙型肝炎病毒增强剂ⅡB1结合因子(hB1F),并将其鉴定为Ftz-F1(NR5A)核受体亚家族的新成员。尽管最近已取得进展,但其生物学功能仍未确定。这项研究的目的是建立hBlF转基因小鼠模型,以促进hB1F的功能研究。方法:将转基因片段显微注射到小鼠受精卵中。被操纵的胚胎被转移到假孕雌性小鼠的输卵管中。通过PCR和Southern印迹分析鉴定后代。用RT-PCR和Western印迹分析法分析转基因表达。使用转基因建立者小鼠建立转基因小鼠谱系。通过PCR分析鉴定了F1和F2小鼠。结果:鉴定出7只小鼠带有转基因拷贝。 RT-PCR和蛋白质印迹结果表明,转基因在其中一种转基因小鼠谱系中的心脏,肝,肺,肾和胃中表达。对转基因小鼠的遗传分析表明,转基因在单个位点整合到染色体中,并且稳定地传播。结论:在本研究中,我们建立了hB1F转基因小鼠模型,这将有助于研究hB1F在体内的生物学功能。

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