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人类生长激素基因的5'侧翼序列包含一个细胞特异性控制元件。

The 5'-flanking sequences of the human growth hormone gene contain a cell-specific control element.

作者信息

Cattini P A, Peritz L N, Anderson T R, Baxter J D, Eberhardt N L

出版信息

DNA. 1986 Dec;5(6):503-9. doi: 10.1089/dna.1.1986.5.503.

Abstract

The 5'-flanking sequences of the human growth hormone (hGH) gene contain a cell-specific control element. Hybrid genes containing truncated 5'-flanking DNA fragments from the hGH gene fused to the chloramphenicol acetyl transferase (cat) gene were examined using transient transfection of rat anterior pituitary (GC) and nonpituitary cell lines (HeLa, Rat 2, and KB); preferential expression of these gene hybrids was only observed in GC cells. Deletions through the 5'-flanking sequences of the hGH gene revealed that the region containing nucleotides -230 to -180 is required for efficient cat gene expression in GC cells. This region of DNA is highly homologous to a region of the rGH gene that contains a tissue-specific control element. A hybrid gene containing the tissue-specific control element, but lacking the proximal promoter elements, of the hGH gene upstream from the promoter of the thymidine kinase gene (TKp) from herpes simplex virus ligated to the cat gene produced more CAT activity than the TKp.cat gene in GC cells but not in HeLa cells. These data suggest that the tissue-specific control element located in the 5'-region of the hGH gene can act in the presence of a heterologous promoter and is specific for expression in pituitary cells.

摘要

人类生长激素(hGH)基因的5'侧翼序列包含一个细胞特异性控制元件。使用大鼠垂体前叶(GC)和非垂体细胞系(HeLa、Rat 2和KB)进行瞬时转染,检测了含有来自hGH基因的截短5'侧翼DNA片段与氯霉素乙酰转移酶(cat)基因融合的杂交基因;仅在GC细胞中观察到这些基因杂交体的优先表达。对hGH基因5'侧翼序列进行缺失分析,结果显示GC细胞中cat基因高效表达需要核苷酸-230至-180区域。该DNA区域与rGH基因中包含组织特异性控制元件的区域高度同源。一个含有hGH基因组织特异性控制元件但缺少近端启动子元件的杂交基因,连接到单纯疱疹病毒胸苷激酶基因(TKp)启动子上游的cat基因上,在GC细胞中产生的CAT活性比TKp.cat基因更高,但在HeLa细胞中并非如此。这些数据表明,位于hGH基因5'区域的组织特异性控制元件在存在异源启动子时可发挥作用,且对垂体细胞中的表达具有特异性。

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