Characterization of the Alu-rich 5'-flanking region of the human prothrombin-encoding gene: identification of a positive cis-acting element that regulates liver-specific expression.
Characterization of the Alu-rich 5'-flanking region of the human prothrombin-encoding gene: identification of a positive cis-acting element that regulates liver-specific expression.
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人类凝血酶原编码基因富含 Alu 的 5 侧翼区域的表征：调节肝脏特异性表达的正顺式作用元件的鉴定。
DOI：
10.1016/0378-1119(90)90368-2
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发表时间：
1990
期刊：
影响因子：
3.5
通讯作者：
Degen,SJ
中科院分区：
文献类型：
--
作者：
Bancroft,JD;Schaefer,LA;Degen,SJ
测定了人凝血酶原编码基因（F2）上游6127 bp的核苷酸序列。由于我们之前表征了转录起始点（tsp）上游417 bp的DNA，因此已知6544 bp的连续侧翼序列。11个Alu重复序列存在于该区域中，占该序列的45%;其他重复序列通过检索GenBank鉴定。通过外显子定位和引物延伸分析，发现tsp具有异质性。为了定位F2肝特异性表达的顺式作用序列，构建了在无启动子的cat基因上游克隆不同长度的F2 5′侧翼区的杂交cat基因。在转染到HepG 2和HeLa细胞中后，推断在HepG 2细胞中合成所需的nt-1101和-798之间的区域;在HeLa细胞中使用这些构建体没有观察到合成。已知的肝脏特异性或调节顺式作用序列的两个序列被确定在该区域。
The nt sequence of 6127 bp of sequence upstream of the human prothrombin-encoding gene (F2) has been determined. Since we previously characterized 417 bp of DNA immediately upstream from the transcription start point (tsp), 6544 bp of continuous flanking sequence are known. Eleven Alu repeat sequences present in this region comprise 45% of the sequence; other repetitive sequences were identified by searching GenBank. The tsp was found to be heterogeneous by exon mapping and primer extension analysis. To localize the cis-acting sequences responsible for the liver-specific expression of F2, hybrid cat genes were constructed with various lengths of F2 5′-flanking region cloned upstream from a promoterless cat gene. After transfection into HepG2 and HeLa cells, it was inferred that the region between nt− 1101 and− 798 was required for synthesis in HepG2 cells; no synthesis was observed using these constructs in HeLa cells. Two sequences for known liver-specific or regulatory cis-acting sequences were identified in this region.