人肺成纤维细胞系WI-38表达多种磷酸酪氨酸磷酸酶mrna。

Receptors & signal transduction Pub Date : 1997-01-01
M A Dayton, T J Knobloch
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引用次数: 0

摘要

蛋白酪氨酸磷酸酶是信号转导途径的重要组成部分。作者利用逆转录/聚合酶链反应完成了58种不同哺乳动物蛋白酪氨酸和双特异性磷酸酶(PTPase)和PTPase样基因在正常人类二倍体成纤维细胞系WI-38中的RNA表达的全面检查。这些PTPase基因中有37个表达易于测量的RNA,其中4个同时表达两种或两种以上同种异构体的RNA。另外8个PTPase基因的信息在低水平时可以检测到。在我们的条件下,只有14个已知的PTPase基因不表达可测量的RNA。为了进行比较,作者还使用高度简并的引物对经典酪氨酸特异性PTPases中发现的保守基序进行了评估,以评估WI-38细胞系中表达的PTPases。使用特异引物检测到的22个经典酪氨酸特异性PTPases中,只有8个被这些简并引物检测到。我们的特异性PTPase引物小组对于半定量评估细胞表达的PTPase库非常有用。
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Multiple phosphotyrosine phosphatase mRNAs are expressed in the human lung fibroblast cell line WI-38.

Protein tyrosine phosphatases are important components of signal transduction pathways. The authors have used reverse transcription/polymerase chain reactions to accomplish a comprehensive examination of the RNA expression for 58 distinct mammalian protein tyrosine and dual specificity phosphatase (PTPase) and PTPase-like genes in the normal human diploid fibroblast cell line WI-38. Thirty-seven of these PTPase genes express easily measurable RNA, and four simultaneously express the RNA for two or more isoforms. Messages for an additional eight PTPase genes are detectable at low levels. Only 14 known PTPase genes do not express measurable RNA under our conditions. For purposes of comparison, the authors also assessed the PTPases expressed in the WI-38 cell line using highly degenerate primers to conserved motifs found in the classical tyrosine-specific PTPases. Only eight of the 22 classic tyrosine-specific PTPases detected using the specific primers were detected using these degenerate primers. Our panel of specific PTPase primers should be very useful for semiquantitatively assessing the repertoire of PTPases expressed by cells.

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