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激肽B(2)受体基因结构、产物加工及在成年和胎鼠中的表达:基因进化的证据

The kinin B(2) receptor gene structure, product processing and expression in adult and fetal rats: evidence for gene evolution.

作者信息

França C E, Vicari C F, Piza A M, Geroldo E A, Beçak M L, Beçak W, Stocco R C, Lindsey C J

机构信息

Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brasil.

出版信息

Genet Mol Res. 2010 Feb 9;9(1):215-30. doi: 10.4238/vol9-1gmr681.

Abstract

We examined the structure of the rat kinin B2 receptor gene (KB2r) and encoding messenger RNA (mRNA) processing. Differently from the closely related mouse and rabbit genes that have three exons and two introns, the rat gene purportedly consists of four exons and three introns. There are two purported gene products; one of them contains an upstream approximately 180-bp open reading frame region ("exon-X") potentially expressed as a result of alternative processing. To examine the processing of rat KB2r mRNA, cDNA amplicons were generated using primer pairs directed towards 5' or 3' exon or intron flanking regions. Analyses of intron/exon primary cDNA amplicons showed that introns 1 to 3 are removed sequentially and that "exon-X" removal follows that of intron-3. No evidence was found for "exon-X" expression in polyadenylated (mature) mRNA of adult Wistar, Wistar Kyoto, spontaneously hypertensive or Sprague-Dawley rat tissues. Nor was "exon-X" detected in tissues subject to inflammatory stimulus expressing B1 kinin receptor mRNA or in 1- to 21-day-old rat embryos or fetuses. The lack of evidence for the expression of "exon-X" in mature mRNA indicates that the structure of the rat gene is similar to that of the mouse, rabbit and human genes, all consisting of three exons and two introns. The "exon-X" fragment may result from interstitial gene duplication, be a fragment of the ancestral gene, or most likely heterologous transposon insertion of an exon-like fragment into intron-2 of the KB2r gene.

摘要

我们研究了大鼠缓激肽B2受体基因(KB2r)的结构及其编码信使核糖核酸(mRNA)的加工过程。与密切相关的小鼠和兔基因不同,后者有三个外显子和两个内含子,大鼠基因据称由四个外显子和三个内含子组成。有两种据称的基因产物;其中一种含有一个上游约180碱基对的开放阅读框区域(“外显子X”),可能由于可变加工而表达。为了研究大鼠KB2r mRNA的加工过程,使用针对5'或3'外显子或内含子侧翼区域的引物对生成了cDNA扩增子。对内含子/外显子初级cDNA扩增子的分析表明,内含子1至3是依次去除的,“外显子X”的去除发生在内含子3之后。在成年Wistar、Wistar Kyoto、自发性高血压或Sprague-Dawley大鼠组织的多聚腺苷酸化(成熟)mRNA中未发现“外显子X”表达的证据。在表达B1缓激肽受体mRNA的炎症刺激组织中,或在1至21日龄大鼠胚胎或胎儿中也未检测到“外显子X”。成熟mRNA中缺乏“外显子X”表达的证据表明,大鼠基因的结构与小鼠、兔和人类基因相似,均由三个外显子和两个内含子组成。“外显子X”片段可能是由于基因间重复产生的,是祖先基因的一个片段,或者最有可能是一个类似外显子的片段异源转座插入到KB2r基因的内含子2中。

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