DNA sequences responsible for tissue‐specific expression of a chicken alpha‐crystallin gene in mouse lens cells.
DNA sequences responsible for tissue‐specific expression of a chicken alpha‐crystallin gene in mouse lens cells.
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负责小鼠晶状体细胞中鸡 α-晶状体蛋白基因组织特异性表达的 DNA 序列。
DOI：
10.1002/j.1460-2075.1985.tb03975.x
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
T. Okada
中科院分区：
文献类型：
--
作者：
K. Okazaki;K. Yasuda;H. Kondoh;T. Okada
我们通过将杂交α/δ晶状体蛋白基因导入原代培养的小鼠透镜上皮细胞的细胞核中，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。使用抗δ-晶状体蛋白血清，通过Western印迹分析确定由α-晶状体蛋白基因的5′上游启动子区与δ-晶状体蛋白基因的结构部分融合组成的杂合基因的瞬时表达水平。杂交基因似乎以组织特异性方式表达，因为它在小鼠透镜细胞中有活性，但在成纤维细胞或L细胞中无活性。位于转录起始位点上游242 - 189 bp的DNA序列是在透镜细胞中高水平表达所必需的。当它们的方向在原始位点反转时，或者当它们位于杂交基因的第二内含子中的帽位点下游约1.7 kbp时，它们是有活性的。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，杂交基因在透镜细胞和成纤维细胞中表达。
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.