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首页> 外文期刊>The Journal of Reproduction and Development >Molecular Cloning of Rat and Porcine Retina-derived POU Domain Factor 1 (POU6F2) from a Pituitary cDNA Library
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Molecular Cloning of Rat and Porcine Retina-derived POU Domain Factor 1 (POU6F2) from a Pituitary cDNA Library

机译:大鼠和猪视网膜来源的POU域因子1(POU6F2)从垂体cDNA文库的分子克隆

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Homeobox transcription factors are known to play crucial roles in the anterior lobe of the pituitary gland. During molecular cloning with the Yeast One-Hybrid System using a 5'-upstream region of the porcine Fsh beta as a bait sequence, we have cloned a cDNA encoding a partial sequence of the retina-derived POU domain factor 1 (RPF1) from the porcine pituitary cDNA library and confirmed its specific binding to the bait sequence. In situ hybridization was performed to examine localization of Rpf1 and showed that this gene is expressed in the stem/progenitor cells of the rat pituitary primordium as well as the diencephalon and retina. In addition, real-time PCR demonstrated that Rpf1 transcripts are abundant in early embryonic periods but that this is followed by a decrease during pituitary development, indicating that this factor plays a role in differentiating cells of the pituitary. The transcriptional activity of RPF1 for genes of Prop1, Prrx1 and Prrx2, which were characterized as genes participating in the pituitary stem/progenitor cells by our group, was then examined with full-length cDNA obtained from the rat pituitary. RPF1 showed regulatory activity for Prop1 and Prrx2, but not for Prrx1. These results indicate the involvement of this retina-derived factor in pituitary development
机译:同源异型盒转录因子在垂体前叶中起着至关重要的作用。在使用酵母Fsh beta的5'上游区域作为诱饵序列的酵母单杂交系统进行分子克隆的过程中,我们已经从cDNA克隆了一个编码部分视网膜衍生POU域因子1(RPF1)序列的cDNA。猪垂体cDNA文库,并证实其与诱饵序列特异性结合。进行了原位杂交以检查Rpf1的定位,并表明该基因在大鼠垂体原基的干/祖细胞以及间脑和视网膜中表达。此外,实时PCR证实Rpf1转录本在早期胚胎期很丰富,但随后在垂体发育过程中下降,这表明该因子在分化垂体细胞中起作用。然后用从大鼠垂体获得的全长cDNA检查RPF1对Prop1,Prrx1和Prrx2基因的转录活性,这些基因被我们的研究组认为是参与垂体干/祖细胞的基因。 RPF1对Prop1和Prrx2显示调节活性,但对Prrx1没有显示调节活性。这些结果表明该视网膜源因子参与垂体发育

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