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大鼠含同源框序列的克隆与表达

Cloning and expression of rat homeo-box-containing sequences.

作者信息

Falzon M, Sanderson N, Chung S Y

出版信息

Gene. 1987;54(1):23-32. doi: 10.1016/0378-1119(87)90343-x.

Abstract

Six rat homeo-box-containing DNA sequences have been isolated by screening a genomic library with a probe derived from an Antennapedia (Antp) cDNA clone of Drosophila melanogaster. Sequence determination of two of the clones containing the homeo-box regions reveals a 180-nucleotide(nt) domain sharing more than 80% homology at the nucleotide level and more than 90% homology at the amino acid level with the homeo-box from the Antp gene and from homeo-boxes of other metazoan species. Genomic blotting experiments suggest that the two homeo-box-containing DNA regions are present in one or two copies per haploid rat genome. Northern blot analysis of RNA has shown that the rat homeo-box sequences are expressed in a tissue-specific manner; transcripts were detected in the spinal cord and kidney, but not in brain, testis, liver, and spleen. The rat nucleotide sequences lying outside the 180-nt homeo-box domain share virtually no sequence homology with the Antp flanking regions. However, one clone does show an equally high degree of amino acid homology within the homeo-box and its immediate flanking region with a putative homologous gene in mouse. The result suggests that some of the mammalian homeo-box-containing genes are conserved in evolution and may serve important cellular or developmental functions.

摘要

通过用源自黑腹果蝇触角足基因(Antp)cDNA克隆的探针筛选基因组文库,已分离出六个含有大鼠同源异型框的DNA序列。对其中两个含有同源异型框区域的克隆进行序列测定,结果显示一个180个核苷酸(nt)的结构域,在核苷酸水平上与Antp基因以及其他后生动物物种的同源异型框具有80%以上的同源性,在氨基酸水平上具有90%以上的同源性。基因组印迹实验表明,每个单倍体大鼠基因组中,这两个含有同源异型框的DNA区域以一或两个拷贝形式存在。对RNA的Northern印迹分析表明,大鼠同源异型框序列以组织特异性方式表达;在脊髓和肾脏中检测到转录本,但在脑、睾丸、肝脏和脾脏中未检测到。位于180 nt同源异型框结构域之外的大鼠核苷酸序列与Antp侧翼区域几乎没有序列同源性。然而,一个克隆在同源异型框及其紧邻的侧翼区域内,与小鼠中的一个假定同源基因显示出同样高度的氨基酸同源性。结果表明,一些含哺乳动物同源异型框的基因在进化过程中是保守的,可能具有重要的细胞或发育功能。

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