人Ⅰ型胶原基因第一内含子调节转录的研究
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Study on the First Intron Regulating Transcription of Human α1(Ⅰ) Collagen Gene
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    摘要:

    人Ⅰ型胶原α1(Ⅰ)链(COLⅠA1)基因内含子序列在不同细胞内有不同的转录调节活性.报道了含人COLⅠA1基因内含子Ⅰ不同区段(+544~+855和+820~+1 093)的重组质粒pSCEP-CAT和pSCIP-CAT的构建并转染人胚肌腱成纤维细胞和Tca 8113舌癌细胞.地高辛标记抗CAT-ELISA检测结果显示:pSCEP-CAT在两种细胞均获表达;pSCIP-CAT在人成纤维细胞未表达,但在舌癌细胞其表达量明显高于pSCEP-CAT.这表明人COLⅠA1基因内含子Ⅰ+544~+855区段增强氯霉素乙酰转移酶(CAT)表达;+820~+1093区段则抑制人成纤维细胞但显著增强舌癌细胞的CAT表达.

    Abstract:

    The first intron sequence of human α1(Ⅰ) collagen (COLⅠA1) gene has been shown to have differential activities of transcription regulation in a variety of tissue cells. To determine the activity of the first intron of COLⅠA1 gene and then utilize the regulatory elements to perform ectopic expression of foreign genes in human fibroblasts. Two novel recombinants pSCEP-CAT and pSCIP-CAT were constructed in which the intronic +544~+855 and +820~+1 093 segment of COLⅠA1 gene were cloned respectively in the upstream position of the promoter sequence of this gene followed by chloramphenical acetyltransferases (CAT) gene. Human fetal tendon fibroblasts and Tca 8113 tongue cancer cells in culture were transfected with the recombinants by using LipofectAMINETM, and then tested the expression level of CAT gene with DIG-labeled anti-CAT ELISA. The results indicate that the first intronic +544~+855 segment of COLⅠA1 gene has an enhancing effect on the expression of CAT gene driven by the promoter in the tranfected human fetal tendon fibroblasts and Tca 8113 tongue cancer cells. Interestingly, the first intronic +820~+1 093 segment has strongly stimulating effect on the expression of the marker gene in the tongue cancer cells, but inhibitory effect in the fibroblasts.

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陈俊杰,杨绍华,王若菡,彭文珍,马英红,刘智敏,李昌龙.人Ⅰ型胶原基因第一内含子调节转录的研究[J].生物化学与生物物理进展,1998,25(4):350-354

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  • 收稿日期:1997-05-02
  • 最后修改日期:1997-10-30
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