Bräuer C, Hanes J, Scheit K H
Max Planck Institute for Biophysical Chemistry, Department of Molecular Biology, Göttingen, Germany.
Biol Chem Hoppe Seyler. 1995 Oct;376(10):631-6.
We have characterized the gene of SVSP109, specifying the bovine secretory protein SVSP109, which is synthesized in a tissue- as well as species-specific manner. Approximately 1.3 kb upstream of the SVSP109 gene, the 3'-end of another gene designated HG5 was identified. The HG5 gene fragment comprises exon (n-1) and exon (n), separated by an intron. Exon (n) contains the complete 3'-untranslated region, whereas exon (n-1) encodes the C-terminal part of a hitherto unknown protein with high homology to SVSP109. The sequenced region of 6956 bp of a bovine genomic clone comprised the complete SVSP109 gene, which is made up of five exons and four introns. Primer extension analysis and RTPCR of poly(A+)RNA from seminal vesicle revealed that the first exon 1 extends to a position 34 bp downstream of the TATA sequence. Employing a CAT assay, a definitive but weak promoter activity was detected in pCATeSVSP15 (base pairs -639 to +574) and pCATeSVSP10 (base pairs -639 to +198); pCATeSVSP6 (base pairs -262 to +65) displayed promoter activity similar to the positive control. We conclude from these results that the TATA sequence located at position -34 is part of the functional promoter of the SVSP109 gene.
我们已经对SVSP109基因进行了特征分析,该基因编码牛分泌蛋白SVSP109,其合成具有组织特异性和物种特异性。在SVSP109基因上游约1.3 kb处,鉴定出另一个名为HG5的基因的3'末端。HG5基因片段包含外显子(n-1)和外显子(n),由一个内含子隔开。外显子(n)包含完整的3'非翻译区,而外显子(n-1)编码一种迄今未知的与SVSP109具有高度同源性的蛋白质的C末端部分。一个牛基因组克隆的6956 bp测序区域包含完整的SVSP109基因,该基因由五个外显子和四个内含子组成。来自精囊的poly(A+)RNA的引物延伸分析和RT-PCR显示,第一个外显子1延伸到TATA序列下游34 bp的位置。采用CAT分析,在pCATeSVSP15(碱基对-639至+574)和pCATeSVSP10(碱基对-639至+198)中检测到明确但较弱的启动子活性;pCATeSVSP6(碱基对-262至+65)显示出与阳性对照相似的启动子活性。我们从这些结果得出结论,位于-34位置的TATA序列是SVSP109基因功能启动子的一部分。