Donoviel D B, Framson P, Eldridge C F, Cooke M, Kobayashi S, Bornstein P
Department of Biochemistry, University of Washington, Seattle 98195.
J Biol Chem. 1988 Dec 15;263(35):18590-3.
We have isolated a 34-kilobase pair genomic thrombospondin clone and determined the sequence of 2033 base pairs of the 5'-flanking region, the first entirely untranslated exon and the first intron. A number of interesting regulatory motifs were identified. The transcription start site, determined by primer extension and S1 nuclease analysis, was shown to be 20-30 bases 3' of a TATA-like sequence, TTTAAAA. We have also shown that a thrombospondin promoter-bovine growth hormone fusion gene is transcribed in transiently transfected cells. We conclude that the genomic clone contains the transcriptional signals of the thrombospondin gene and will therefore be useful in the analysis of cis-acting elements that regulate the expression of this gene.
我们分离出了一个34千碱基对的基因组血小板反应蛋白克隆，并测定了其5'侧翼区2033个碱基对、首个完全不翻译的外显子及首个内含子的序列。鉴定出了一些有趣的调控基序。通过引物延伸和S1核酸酶分析确定的转录起始位点，位于类似TATA的序列TTTAAAA下游20 - 30个碱基处。我们还表明，血小板反应蛋白启动子 - 牛生长激素融合基因在瞬时转染细胞中能够转录。我们得出结论，该基因组克隆包含血小板反应蛋白基因的转录信号，因此将有助于分析调控该基因表达的顺式作用元件。
