Genomic structure and promoter activity of the human tissue factor pathway inhibitor-2 gene
Genomic structure and promoter activity of the human tissue factor pathway inhibitor-2 gene
复制标题
DOI：
10.1016/s0167-4781(00)00298-0
复制
发表时间：
2001-02-16
期刊：
影响因子：
--
通讯作者：
Kisiel, W
中科院分区：
文献类型：
--
作者：
Kamei, S;Kazama, Y;Kisiel, W
人 2 型组织因子途径抑制剂 (TFPI-2)，也称为胎盘蛋白 5，是一种 32 kDa 丝氨酸蛋白酶抑制剂，由三个串联排列的 Kunitz 型抑制剂结构域组成，与组织因子途径抑制剂同源。 TFPI-2 强烈抑制多种丝氨酸蛋白酶，包括胰蛋白酶、胰凝乳蛋白酶、纤溶酶、激肽释放酶和凝血因子 XIa。在这项研究中，我们从人工染色体基因组文库中分离并表征了一个基因组克隆，该克隆编码整个人类 TFPI-2 基因。人类 TFPI-2 基因长约 7 kb，由五个外显子和四个内含子组成。每个 Kunitz 型结构域均由单个外显子编码，类似于鼠 TFPI-2 和其他 Kunitz 型蛋白酶抑制剂中观察到的情况。总共 535 bp 的 3' 侧翼区域包含两个可能的聚腺苷酸化位点 (AATAAA)，位于 +4297 和 +4314。通过寡核苷酸加帽和逆转录 PCR 分析鉴定了单个转录起始位点。将含有 5' 侧翼区域片段的报告质粒瞬时转染至人转化的骨髓内皮细胞和胶质母细胞瘤细胞中，鉴定出足以转录人 TFPI-2 基因的 85 bp 区域（-224 至 -139）。 (C) 2001 Elsevier Science B.V. 保留所有权利。
Human type-2 tissue factor pathway inhibitor (TFPI-2), also known as placental protein 5, is a 32 kDa serine proteinase inhibitor consisting of three tandemly arranged Kunitz-type inhibitor domains homologous to tissue factor pathway inhibitor. TFPI-2 strongly inhibits a wide variety of serine proteinases including trypsin, chymotrypsin, plasmin, kallikrein and blood coagulation factor XIa. In this study, we have isolated and characterized a genomic clone from an artificial chromosome genomic library that encodes the entire human TFPI-2 gene. The human TFPI-2 gene spans approximately 7 kb and consists of five exons and four introns. Each Kunitz-type domain is encoded by a single exon, similar to that observed for murine TFPI-2 and other Kunitz-type proteinase inhibitors. A total of 535 bp of the 3'-flanking region contain two probable polyadenylation sites (AATAAA) at +4297 and +4314. A single transcription initiation site was identified by oligo-capping and reverse transcription-PCR analysis. Transient transfection of reporter plasmids containing segments of the 5'-flanking region into human transformed bone marrow endothelial cells and glioblastoma cells identified an 85 bp region (-224 to -139) sufficient for transcription of the human TFPI-2 gene. (C) 2001 Elsevier Science B.V. All rights reserved.