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Cell-specificity of the chicken ovalbumin and conalbumin promoters.

机译:鸡卵清蛋白和伴清蛋白启动子的细胞特异性。

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摘要

A series of recombinant plasmids containing increasing lengths of the 5'-flanking promoter sequences of the chicken conalbumin and ovalbumin genes fused to the sequences coding for the SV40 T-antigen have been constructed. These recombinants were introduced into a variety of established cell lines and primary cultured cells by nuclear microinjection. Promoter activity was estimated by monitoring T-antigen synthesis by indirect immunofluorescence. We show that the microinjected ovalbumin and conalbumin promoter regions do not function in chicken fibroblasts, kidney cells and in a variety of non-chicken cells, irrespective of the presence of steroid hormone receptors. In contrast, these promoter regions are active in primary cultured chicken embryonic hepatocytes and oviduct tubular gland cells, suggesting the presence of cell-specific transcription factors in these cells. Unexpectedly, promoter sequences close to the TATA boxes of both the ovalbumin and conalbumin genes are sufficient to confer cell-specific expression. Most of the controls exerted on the ovalbumin and conalbumin promoters in the whole animal appear to be reproduced in vitro by nuclear microinjection of the chimeric genes into the primary cultured cells. However, the microinjected ovalbumin promoter is active in embryonic hepatocytes and thus escapes the regulation imposed on the corresponding inactive endogenous gene.
机译:已经构建了一系列重组质粒,其含有与编码SV40 T-抗原的序列融合的鸡伴清蛋白和卵清蛋白基因的5'侧翼启动子序列的长度增加。通过核显微注射将这些重组体引入多种已建立的细胞系和原代培养细胞中。通过通过间接免疫荧光监测T抗原合成来估计启动子活性。我们显示显微注射的卵清蛋白和伴清蛋白启动子区域在鸡成纤维细胞,肾细胞和各种非鸡细胞中均不起作用,无论是否存在类固醇激素受体。相反,这些启动子区域在原代培养的鸡胚肝细胞和输卵管小管腺细胞中具有活性,表明这些细胞中存在细胞特异性转录因子。出乎意料的是,接近卵清蛋白和伴清蛋白基因的TATA框的启动子序列足以赋予细胞特异性表达。整个动物的卵清蛋白和伴清蛋白启动子上的大多数对照似乎是通过将嵌合基因核显微注射到原代培养细胞中而在体外复制的。然而,显微注射的卵清蛋白启动子在胚胎肝细胞中具有活性,因此逃避了对相应的非活性内源基因施加的调节。

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