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牛蛙(Rana catesbeiana)雄激素受体的克隆与鉴定

Cloning and characterization of androgen receptor from bullfrog, Rana catesbeiana.

作者信息

Chattopadhyay Soma, Park Jin Hee, Seong Jae Young, Kwon Hyuk Bang, Lee Keesook

机构信息

Hormone Research Center, Chonnam National University, Gwangju 500-757, South Korea.

出版信息

Gen Comp Endocrinol. 2003 Oct 15;134(1):10-7. doi: 10.1016/s0016-6480(03)00215-6.

Abstract

We have cloned and characterized a full-length cDNA of androgen receptor (AR) from the testis of bullfrog, Rana catesbeiana. The cDNA contains an open reading frame of 2328 nucleotides encoding a protein of 776 amino acid residues. The bullfrog AR shows high homology with ARs from other species in its amino acid sequence. Its overall homology with those of African clawed frog, Japanese eel and human is 70, 53, and 63%, respectively. As expected, the N-terminal domain shows much less homology (30-59%) than both DNA-binding domain (85-92%) and ligand binding domain (80-89%). Northern blot analysis detected the bullfrog AR message as a single transcript of around 9 kb only in the testis. However, RT-PCR analysis revealed that AR mRNA is also expressed in other tissues although the levels are very low compared to that in the testis. Western blot analysis of whole tissue extracts showed the presence of AR protein in fore brain, heart, and testis. The AR gene is present as a single copy in bullfrog based on Southern blot analysis of genomic DNA. Altogether, the results suggest that the bullfrog AR is evolutionary conserved and may have functions similar to those shown in other species.

摘要

我们已经从牛蛙(Rana catesbeiana)的睾丸中克隆并鉴定了雄激素受体(AR)的全长cDNA。该cDNA包含一个2328个核苷酸的开放阅读框,编码一个由776个氨基酸残基组成的蛋白质。牛蛙AR在氨基酸序列上与其他物种的AR具有高度同源性。它与非洲爪蟾、日本鳗鲡和人类的AR的总体同源性分别为70%、53%和63%。正如预期的那样,N端结构域的同源性(30 - 59%)远低于DNA结合结构域(85 - 92%)和配体结合结构域(80 - 89%)。Northern印迹分析仅在睾丸中检测到牛蛙AR信息为一条约9 kb的单一转录本。然而,RT-PCR分析表明,尽管与睾丸中的水平相比非常低,但AR mRNA也在其他组织中表达。对全组织提取物的Western印迹分析显示,在前脑、心脏和睾丸中存在AR蛋白。基于基因组DNA的Southern印迹分析,AR基因在牛蛙中以单拷贝形式存在。总之,结果表明牛蛙AR在进化上是保守的,可能具有与其他物种中所示类似的功能。

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