Identification of a cis-acting element that enhances the pigment cell-specific expression of the human tyrosinase gene.
Identification of a cis-acting element that enhances the pigment cell-specific expression of the human tyrosinase gene.
复制标题
鉴定增强人酪氨酸酶基因色素细胞特异性表达的顺式作用元件。
DOI：
10.1016/s0021-9258(19)36726-2
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
S. Shibahara
中科院分区：
文献类型：
--
作者：
K. Shibata;Y. Muraosa;Y. Tomita;Hachiro Tagami;S. Shibahara
为了鉴定负责人酪氨酸酶基因色素细胞特异性表达的顺式作用元件，我们通过瞬时表达测定分析了其5 '侧翼区的启动子活性。通过在萤火虫荧光素酶基因上游插入人酪氨酸酶基因的5 ′侧翼区构建融合基因，并将其导入人黑素瘤细胞和HeLa细胞。因此，我们发现的元素，位于2.7和1.8之间的转录起始位点上游的酪氨酸酶对，增强荧光素酶报告基因在黑色素瘤细胞中的瞬时表达，但不是在HeLa细胞，酪氨酸酶基因的表达是不可检测的。使用含有在异源猿猴病毒40启动子控制下的推定增强子元件的融合基因，我们鉴定了在-2.0和-1.8个酶对之间的约200个碱基对（bp）的色素细胞特异性增强子，并将核心序列定位于39 bp区域。然后证实该39 bp的核心元件指导酪氨酸酶基因启动子下的报告基因的黑色素瘤细胞特异性表达。因此，我们建议，这个核心元素是负责色素细胞特异性表达的人酪氨酸酶基因。
To identify the cis-acting element that is responsible for the pigment cell-specific expression of the human tyrosinase gene, we analyzed the promoter activity of its 5‘-flanking region by transient expression assays. The fusion genes were constructed by inserting the 5‘-flanking region of the human tyrosinase gene upstream from the firefly luciferase gene and were introduced into human melanoma cells and HeLa cells. We thus found the element, located between 2.7 and 1.8 kilobase pairs upstream from the transcription initiation site, that enhances the transient expression of the luciferase reporter gene in melanoma cells, but not in HeLa cells, the tyrosinase gene expression of which is not detectable. Using the fusion genes containing putative enhancer elements under the control of the heterologous simian virus 40 promoter, we identified the pigment cell-specific enhancer of approximately 200 base pairs (bp) between -2.0 and -1.8 kilobase pairs and localized the core sequence to a 39-bp region. This 39-bp core element was then confirmed to direct the melanoma cell-specific expression of the reporter gene under the tyrosinase gene promoter. We thus propose that this core element is responsible for the pigment cell-specific expression of the human tyrosinase gene.