牛精液主要蛋白(SVSP109)基因:结构与启动子分析。

Biological chemistry Hoppe-Seyler Pub Date : 1995-10-01
C Bräuer, J Hanes, K H Scheit
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引用次数: 0

摘要

我们已经鉴定了SVSP109的基因,确定了牛分泌蛋白SVSP109,该蛋白以组织特异性和物种特异性的方式合成。在SVSP109基因上游约1.3 kb处,发现了另一个名为HG5的基因的3'端。HG5基因片段包括外显子(n-1)和外显子(n),由内含子分隔。外显子(n)包含完整的3'-未翻译区,而外显子(n-1)编码一种迄今未知的蛋白质的c端部分,与SVSP109具有高度同源性。牛基因组克隆的6956 bp序列区包含完整的SVSP109基因,该基因由5个外显子和4个内含子组成。引物延伸分析和精囊poly(A+)RNA的RTPCR显示,第一个外显子1延伸到TATA序列下游34bp的位置。采用CAT检测,在pCATeSVSP15(碱基对-639至+574)和pCATeSVSP10(碱基对-639至+198)中检测到明确但微弱的启动子活性;pCATeSVSP6(碱基对-262 ~ +65)的启动子活性与阳性对照相似。我们从这些结果中得出结论,位于-34位的TATA序列是SVSP109基因功能启动子的一部分。
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The gene for the major protein (SVSP109) of bovine semen: structure and promoter analysis.

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.

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