Ran结合蛋白RanBP1的同源基因M3的克隆与功能分析

Cloning and Function Analysis of M3: A Gene Homologous to Ran Binding Protein RanBP1

以Ran结合蛋白RanBP1的保守区域作为探针 ,低严谨条件下杂交筛选人视网膜cDNA文库 ,得到阳性克隆M 3,它与鼠zhx 1基因高度同源 .RH作图将其定位于 8q2 4.11～q2 4.3.拼接EST序列并填补缺口 ,得到M 3cDNA全长 4934bp ,与Northernblot得到的主要转录本长度一致 .此cDNA全序列包括 2 6 2 2bp开放阅读框 ,编码 873氨基酸 ,含有 2种翻译调控元件uUAG和TTATTTAT .该基因预测蛋白含有 2个锌指结构 ,5个同源盒保守区 ,1个双侧核定位序列 (NLS)及 35个磷酸化位点 ,表明它既可能作为转录调控因子 ,也可能受到上游因子的调控 .

The conserved region of Ran Binding Protein RanBP1 was used as a probe to screen the human retina cDNA library under low stringency. A positive clone named M3 was obtained, which was highly homologous to zhx 1 gene in M. musculus. The gene was positioned to the region of 8q24.11 q24.3 by RH mapping. The full length of the M3 cDNA(obtained by EST assembling)was 4 934 bp, corresponding to that of the main transcript shown in Northern bolt. This cDNA sequence included an open reading frame of 2 622 bp(coding for 873 aa) and two kinds of translation modules uAUG and TTATTTAT. The predicted protein had two C 2H 2 zinc finger structures, five homeodomains, one bipartite nuclear localization signal(NLS) and 35 potential phosphorylation sites, which all suggested that M3 gene may both function as transcriptional regulator and be regulated by other upstream factors.