Casalotti S O, Pelaia G, Yakovlev A G, Csikós T, Grayson D R, Krueger K E
Fidia-Georgetown Institute for the Neurosciences, Georgetown University School of Medicine, Washington, DC 20007.
Gene. 1992 Nov 16;121(2):377-82. doi: 10.1016/0378-1119(92)90147-h.
The gene encoding the rat mitochondrial benzodiazepine receptor (MBR) was cloned and characterized. Hybridization of a previously cloned cDNA for MBR to genomic Southern blots indicated that the gene was probably present at one copy per haploid genome. Rapid amplification of cDNA ends with rat adrenal RNA was used to obtain 47 nt of additional sequence upstream from our previously cloned MBR cDNA proving to be a crucial step in cloning the first exon of this gene. The MBR gene is comprised of four exons spanning approx. 10 kb. The first intron, contained within a 8-kb stretch of this gene, is located within the 5'-untranslated sequence, whereas the remaining two introns are much shorter (641 and 854 bp) and interrupt the coding sequence. The third intron contains sequences homologous to rodent B1 repetitive elements and a novel sequence closely resembling part of a repetitive element belonging to the Alu family in humans. The transcription start point was mapped by S1 nuclease protection assays suggesting that the first exon is just 56 bp in length. The sequence upstream from this region contains three GC boxes but lacks other known consensus recognition sites for sequence-specific transcription factors.
编码大鼠线粒体苯二氮䓬受体(MBR)的基因被克隆并进行了特征分析。先前克隆的MBR cDNA与基因组Southern杂交结果表明,该基因可能在单倍体基因组中以单拷贝形式存在。利用大鼠肾上腺RNA进行cDNA末端快速扩增,获得了先前克隆的MBR cDNA上游47个核苷酸的额外序列,这被证明是克隆该基因第一个外显子的关键步骤。MBR基因由四个外显子组成,跨度约为10 kb。该基因8 kb区域内的第一个内含子位于5'-非翻译序列中,而其余两个内含子则短得多(分别为641和854 bp),并打断编码序列。第三个内含子包含与啮齿动物B1重复元件同源的序列以及一个与人类Alu家族重复元件部分非常相似的新序列。通过S1核酸酶保护试验确定了转录起始点,表明第一个外显子长度仅为56 bp。该区域上游的序列包含三个GC框,但缺乏序列特异性转录因子的其他已知共有识别位点。