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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Rod photoreceptor-specific gene expression in human retinoblastoma cells.
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Rod photoreceptor-specific gene expression in human retinoblastoma cells.

机译：杆状感光细胞特异性基因在人视网膜母细胞瘤细胞中的表达。

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摘要

Retinoblastoma cells in culture have previously been shown to express cone-specific genes but not their rod counterparts. We have detected the messages for the rod alpha, beta, and gamma subunits of cGMP phosphodiesterase (PDE), the rod alpha subunit of transducin, rod opsin, and the cone alpha' subunit of PDE in RNA of human Y-79 retinoblastoma cells by reverse transcription-PCR. Quantitative analysis of the mRNAs for the rod alpha and cone alpha' PDE subunits revealed that they were expressed at comparable levels; however, the transcript encoding the rod beta PDE subunit was 10 times more abundant in these cells. Northern hybridization analysis of Y-79 cell RNA confirmed the presence of the transcripts for rod and cone PDE catalytic subunits. To test whether the transcriptional machinery required for the expression of rod-specific genes was endogenous in Y-79 retinoblastoma cells, cultures were transfected with a construct containing the promoter region of the rod beta PDE subunit gene attached to the firefly luciferase reporter vector. Significant levels of reporter enzyme activity were observed in the cell lysates. Our results demonstrate that the Y-79 retinoblastoma cell line is a good model system for the study of transcriptional regulation of rod-specific genes.
机译：以前已证明培养物中的成视网膜细胞瘤细胞表达视锥细胞特异性基因，但不表达其视杆对应物。我们已经通过以下方法检测到了人G-磷酸二酯酶（PDE）的杆状α，β和γ亚基，转导素，杆视蛋白的杆状α亚基以及人Y-79视网膜母细胞瘤细胞RNA中PDE的锥状α'亚基的信息。逆转录PCR。杆α和锥α'PDE亚基的mRNA的定量分析表明，它们的表达水平相当。然而，编码杆βPDE亚基的转录本在这些细胞中的含量是后者的10倍。 Y-79细胞RNA的Northern杂交分析证实了棒状和锥状PDE催化亚基的转录本的存在。为了测试表达杆特异性基因所需的转录机制在Y-79视网膜母细胞瘤细胞中是否是内源的，将培养物转染到含有萤火虫荧光素酶报告载体的杆βPDE亚基基因启动子区域的构建体上。在细胞裂解物中观察到显着水平的报道酶活性。我们的结果表明，Y-79视网膜母细胞瘤细胞系是研究杆特异性基因转录调控的良好模型系统。

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