Staubli F, Klein A, Rentsch J M, Hameister H, Berchtold M W
Institute of Veterinary Biochemistry, University of Zurich-Irchel. Switzerland.
Mamm Genome. 1995 Nov;6(11):769-77. doi: 10.1007/BF00539001.
The rat gene encoding oncomodulin (OM), a small calcium-binding protein, is under the control of a solo LTR derived from an endogenous intracisternal A-particle. The latter sequence is the only OM promoter analyzed so far. In order to study cell type-specific OM expression in a species lacking LTR sequences in the OM locus, we initially synthesized an OM cDNA from mouse placenta. By sequencing, we found a 137-bp-long 5'leader region that differed markedly from its rat counterpart but had high similarity to several mouse genomic sequences. Primers specific to this sequence in addition with primers specific for an exon 2/intron 2 sequence were used to screen a mouse ES cell line genomic P1 library. One positive clone contained the whole OM gene, including intron 1 of 25kb and a 5' flanking region of 27 kb lacking an LTR. The region upstream of exon 1 contains no TATA or CCAAT boxes but has a homopurine/homopyrimidine stretch of 102 bp as well as a (CA)22 repeat. The latter sequence is polymorphic and was therefore, used to map the OM gene to the distal end of the long arm of mouse Chromosome (Chr) 5 by interspecific backcross analysis. Additionally we localized the OM gene by in situ hybridization to the region G1-3 on Chr 5, confirming the genetic linkage results. Finally, the OM gene was found to be structurally conserved and to exist in a single copy in mammals.
编码癌调蛋白(OM)的大鼠基因是一种小的钙结合蛋白,受源自内源性脑池内A颗粒的单个长末端重复序列(LTR)调控。后一序列是迄今为止分析的唯一OM启动子。为了研究在OM基因座中缺乏LTR序列的物种中细胞类型特异性OM表达,我们最初从小鼠胎盘中合成了OM cDNA。通过测序,我们发现了一个137bp长的5'前导区,它与大鼠对应区域明显不同,但与几个小鼠基因组序列高度相似。除了针对外显子2/内含子2序列的引物外,还使用针对该序列的特异性引物筛选小鼠胚胎干细胞系基因组P1文库。一个阳性克隆包含整个OM基因,包括25kb的内含子1和27kb的5'侧翼区域,该区域缺乏LTR。外显子1上游区域没有TATA或CCAAT框,但有一个102bp的同型嘌呤/同型嘧啶延伸以及一个(CA)22重复序列。后一序列是多态性的,因此通过种间回交分析用于将OM基因定位到小鼠染色体(Chr)5长臂的远端。此外,我们通过原位杂交将OM基因定位到Chr 5上的G1-3区域,证实了遗传连锁结果。最后,发现OM基因在结构上是保守的,并且在哺乳动物中以单拷贝形式存在。