在Sprague-Dawley大鼠肝脏中表达的两个α - 1b肾上腺素能受体mrna具有不同的5′-区。

C L Deng, L E Cornett
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引用次数: 7

摘要

先前的研究表明,在Sprague Dawley大鼠中,由一个基因转录而来的α - 1b-肾上腺素能受体mrna有两种大小,分别为3.3 kb和2.7 kb,它们在肝脏中的表达量大致相等。只有2.7 kb的mRNA在心脏中表达。两种α - 1b肾上腺素能受体mrna似乎具有广泛的同源区域,因此,我们使用寡核苷酸定向核糖核酸酶H作图来检测两种转录本之间的序列差异。使用寡核苷酸(dT)导向RNase H水解的初步实验表明,这两个mrna具有相同长度的多[A+]尾部。通过区域特异性cDNA探针,我们确定了两个α - 1b肾上腺素能受体mrna的序列差异在于5'端,在2.7 kb转录物中已知的启动物AUG的上游。此外,核糖核酸酶保护实验和使用寡核苷酸作为探针的Northern blot分析结果表明,两种α - 1b-肾上腺素能受体mrna转录自同一条DNA链。
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Two alpha 1b-adrenergic receptor mRNAS expressed in Sprague-Dawley rat liver have distinct 5'-regions.

Previous studies have revealed two size classes of alpha 1b-adrenergic receptor mRNAs, 3.3 kb and 2.7 kb, in the Sprague Dawley rat that are transcribed from a single gene and are expressed in approximately equal amounts in liver. Only the 2.7 kb mRNA is expressed in heart. Both alpha 1b-adrenergic receptor mRNAs appear to share extensive regions of homology, therefore, we used oligonucleotide-directed ribonuclease H mapping to detect sequence differences between the two transcripts. Initial experiments using oligo (dT)-directed RNase H hydrolysis indicated that the two mRNAs have poly [A+] tails of identical length. By using region-specific cDNA probes, we determined that the sequence difference between the two alpha 1b-adrenergic receptor mRNAs lies in the 5' end, upstream from the known initiator AUG in the 2.7 kb transcript. In addition, results from ribonuclease protection assays and Northern blot analysis in which an oligonucleotide was used as the probe suggested that both alpha 1b-adrenergic receptor mRNAs are transcribed from the same DNA strand.

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