Cloning of the Syrian hamster p53 gene: structural and functional characterization of the upstream promoter region.
Cloning of the Syrian hamster p53 gene: structural and functional characterization of the upstream promoter region.
复制标题
叙利亚仓鼠 p53 基因的克隆：上游启动子区域的结构和功能特征。
DOI：
10.1002/mc.2940110309
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发表时间：
1994
影响因子：
4.6
通讯作者：
Notario,V
中科院分区：
文献类型：
--
作者：
Albor,A;Laborda,J;Notario,V
我们用粘粒克隆法从叙利亚仓鼠胚胎细胞中分离到p53基因。仓鼠p53基因的组织结构与其他仓鼠p53基因相似，它有11个外显子，一个非编码外显子1和一个长内含子1（约6.5 kb）。分离上游53启动子，并确定外显子/内含子1边界上游694 bp加上内含子1前6个核苷酸的区域的核苷酸序列。该基因组区域与小鼠和人类的基因组区域高度同源，但包含两个物种中均不存在的重复元件。与鼠和人启动子的序列比较揭示了在431-bp Sacl-Pstl片段内存在相似的转录因子结合基序。原代叙利亚仓鼠胚胎细胞和肿瘤细胞系的瞬时转染试验表明，Sacl-Pstl片段位于细菌氯霉素乙酰转移酶（CAT）基因上游的重组构建体能够有效表达CAT活性。引物延伸分析确定了p53上游启动子内的几个假定的转录起始位点，其中最强的位于外显子/内含子1连接点上游315 bp和包含调控基序的区域上游约30 bp处。©1994 Wiley利斯公司
We isolated thep53gene from Syrian hamster embryo cells by cosmid cloning procedures. The organization of the hamsterp53gene was similar to that of other mammalianp53genes; it had 11 exons, a noncoding exon 1, and a long intron 1 (about 6.5 kb). The upstreamp53promoter was isolated, and the nucleotide sequence of a region encompassing 694 bp upstream from the exon/intron 1 boundary plus the first 6 nt of intron 1 was determined. This genomic region was highly homologous to those of mice and humans but contained a repetitive element not present in either species. Sequence comparisons with the murine and human promoters revealed the presence of similar transcription‐factor binding motifs mapping within a 431‐bp Sacl‐Pstl fragment. Transient transfection assays of primary Syrian hamster embryo cells and neoplastic cell lines with recombinant constructs in which the Sacl‐Pstl fragment was placed upstream of a bacterial chloramphenicol acetyltransferase (CAT) gene revealed the efficient expression of CAT activity. Primer extension analyses identified several putative transcription initiation sites within thep53upstream promoter, the strongest of which was located 315 bp upstream from the exon/intron 1 junction and about 30 bp upstream from the region encompassing the regulatory motifs. ©1994 Wiley‐Liss, Inc.