Epithelial-specific keratin gene expression: identification of a 300 base-pair controlling segment.
Epithelial-specific keratin gene expression: identification of a 300 base-pair controlling segment.
复制标题
上皮特异性角蛋白基因表达：300 个碱基对控制片段的鉴定。
DOI：
10.1093/nar/18.2.247
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发表时间：
1990
影响因子：
14.9
通讯作者：
Blumenberg,M
中科院分区：
文献类型：
--
作者：
Jiang,CK;Epstein,HS;Tomic,M;Freedberg,IM;Blumenberg,M
为了阐明角蛋白在表皮中表达调控所需的元件，我们将一个与人类K#14基因启动子区域相对应的300个碱基对的短片段连接到氯霉素乙酰基转移酶基因。用Ca3(PO4)2沉淀法将其导入多种哺乳动物细胞系和原代培养细胞。角蛋白基因启动子区域的300碱基对片段在所有研究的上皮细胞中都是活性的，包括转化的简单上皮细胞，如HeLa和ME-180，来自复层上皮细胞的细胞系，如SCC-12，以及原代培养的上皮细胞。该结构在所有被测试的非上皮细胞中都是不活跃的，包括成纤维细胞和黑素细胞。该片段在非上皮细胞中不起沉默作用，但在上皮细胞中可以起到增强剂的作用。利用聚合酶链式反应，我们构建了一系列的启动子缺失，并将一个重要的功能定位在一个40bp的序列中。我们的结论是，我们已经确定了角蛋白基因启动子，足以赋予上皮特异性表达。
To elucidate the elements required for regulation of keratin expression in epidermis, we have linked a short, 300 base pair segment, corresponding to the promoter region of a human K # 14 gene, to the chloramphenicolacetyl-transferase gene. This construct was introduced into various mammalian cell lines and primary cultures via Ca3(PO4)2precipitation. The 300 base pair segment from the keratin gene promoter region was active in all epithelial cells studied including transformed, simple epithelial cells such as HeLa and ME-180, cell lines derived from stratified epithelium, such as SCC-12, as well as primary cultures of epithelial cells. The construct was inactive in all non-epithelial cells tested including fibroblasts and melanocytes. The segment does not function as a silencer in nonepithelial cells but it can function as an enhancer in epithelial cells. Using the polymerase chain reaction we have constructed a series of deletions of the promoter and have localized an essential function within a 40 bp sequence. We conclude that we have identified the keratin gene promoter that is sufficient to confer epithelial-specific expression.