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负责鸡α-晶体蛋白基因在小鼠晶状体细胞中组织特异性表达的DNA序列。

DNA sequences responsible for tissue-specific expression of a chicken alpha-crystallin gene in mouse lens cells.

作者信息

Okazaki K, Yasuda K, Kondoh H, Okada T S

出版信息

EMBO J. 1985 Oct;4(10):2589-95. doi: 10.1002/j.1460-2075.1985.tb03975.x.

Abstract

We have studied the DNA sequences required for high-level expression of a cloned chicken alpha-crystallin gene by introducing a hybrid alpha/delta-crystallin gene into nuclei of mouse lens epithelial cells in primary culture. The level of transient expression of the hybrid gene consisting of the 5' upstream promoter region of the alpha-crystallin gene fused to the structural portion of the delta-crystallin gene was determined by Western blot analysis using anti-delta-crystallin serum. The hybrid gene appears to be expressed in a tissue-specific manner, since it is active in mouse lens cells but not in fibroblasts or in L cells. The DNA sequences located 242-189 bp upstream from the transcription initiation site are required for high-level expression in lens cells. They are active when their orientation is reversed at the original site or when placed approximately 1.7 kbp downstream from the cap site in the second intron of the hybrid gene in either orientation. When these DNA sequences were replaced by the enhancer sequences of Moloney murine leukemia virus, the hybrid gene was expressed in both lens cells and fibroblasts.

摘要

我们通过将一个杂交的α/δ-晶状体蛋白基因导入原代培养的小鼠晶状体上皮细胞核中,研究了克隆的鸡α-晶状体蛋白基因高水平表达所需的DNA序列。由α-晶状体蛋白基因的5'上游启动子区域与δ-晶状体蛋白基因的结构部分融合而成的杂交基因的瞬时表达水平,通过使用抗δ-晶状体蛋白血清的蛋白质印迹分析来确定。该杂交基因似乎以组织特异性方式表达,因为它在小鼠晶状体细胞中具有活性,但在成纤维细胞或L细胞中无活性。转录起始位点上游242 - 189 bp处的DNA序列是晶状体细胞中高水平表达所必需的。当它们在原位的方向颠倒时,或者当以任何一种方向置于杂交基因第二个内含子中帽位点下游约1.7 kbp处时,它们都是有活性的。当这些DNA序列被莫洛尼鼠白血病病毒的增强子序列取代时,杂交基因在晶状体细胞和成纤维细胞中均有表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebfa/554548/fbf86be9ab0f/emboj00275-0176-a.jpg

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