通过单步聚合酶链反应测定各种组织中的肾素基因表达。

Renin gene expression in various tissues determined by single-step polymerase chain reaction.

Lou Y K, Smith D L, Robinson B G, Morris B J

Department of Physiology, University of Sydney, New South Wales, Australia.

Clin Exp Pharmacol Physiol. 1991 May;18(5):357-62. doi: 10.1111/j.1440-1681.1991.tb01463.x.

Abstract

Renin mRNA is present in the kidney and, in lower concentrations, in many extrarenal tissues and serves as an index of renin gene activity, as well as potential renin or prorenin synthesis in cell populations within those tissues. Unfortunately the quantity can be very low. 2. A new, highly sensitive technique is described for detection of renin mRNA that involves the enzyme Taq polymerase for both reverse transcription of renin mRNA into renin cDNA and for amplification of the 769-1099 nucleotide segment by the polymerase chain reaction (PCR), all of which involves a single reaction mixture. 3. In this way renin mRNA was detected in kidney and several extrarenal tissues as a PCR product of approximately 330 base pairs on agarose gels by ethidium-bromide staining or hybridization probing. The region of renin mRNA chosen for amplification spanned several intron sites in the coding sequence so that the presence of amplicants derived from genomic DNA could be readily discriminated, as a band of approximately 1.5 kilobases. 4. Thus single-step PCR offers a powerful new approach to detection of renin mRNA.

摘要

肾素信使核糖核酸（Renin mRNA）存在于肾脏中，在许多肾外组织中的浓度较低，它是肾素基因活性的指标，也是这些组织内细胞群体中潜在肾素或肾素原合成的指标。不幸的是，其数量可能非常少。2. 描述了一种用于检测肾素信使核糖核酸的新的高灵敏度技术，该技术涉及Taq聚合酶，用于将肾素信使核糖核酸逆转录为肾素互补脱氧核糖核酸（renin cDNA），并通过聚合酶链反应（PCR）扩增769 - 1099个核苷酸片段，所有这些都在单一反应混合物中进行。3. 通过这种方式，在肾脏和几种肾外组织中检测到肾素信使核糖核酸，作为琼脂糖凝胶上约330个碱基对的PCR产物，通过溴化乙锭染色或杂交探针检测。选择用于扩增的肾素信使核糖核酸区域跨越编码序列中的几个内含子位点，以便能够容易地鉴别出来自基因组脱氧核糖核酸的扩增子，其为一条约1.5千碱基的条带。4. 因此，单步PCR为检测肾素信使核糖核酸提供了一种强大的新方法。