FUNCTIONAL-ANALYSIS OF THE PROMOTER OF A SEA-URCHIN METALLOTHIONEIN GENE
FUNCTIONAL-ANALYSIS OF THE PROMOTER OF A SEA-URCHIN METALLOTHIONEIN GENE
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DOI：
10.1139/o92-160
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发表时间：
1992-10-01
期刊：
影响因子：
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通讯作者：
BRANDHORST, BP
中科院分区：
文献类型：
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作者：
CSERJESI, P;FAIRLEY, P;BRANDHORST, BP
海胆Lytechinus pictus的金属硫蛋白（MT）基因LpMT 1的5 ′-侧翼区包括三个拷贝的保守序列，该保守序列包括重金属诱导其他MT基因所需的金属响应元件（MRE）共有核心序列、GC盒、推定的基础水平增强子元件（包括另一个MRE核心元件）的G盒和poly（C）区。将LpMT 1 DNA的+31 ~-309位核苷酸与氯霉素乙酰转移酶报告基因融合后，注入L. pictus胚胎在仅保留195个碱基对的5 '-侧翼序列的缺失突变体中，这种诱导的活性大大降低，所述缺失突变体包括近端MRE对和G盒，但不包括聚（C）束、GC盒和远端MRE。一个有效的人hMT-II（A）基因启动子在L. pictus胚胎相反，LpMT 1启动子在HeLa细胞和海胆球海胆的胚胎中是活跃的。hMT-II（A）基因可能缺少MT基因在L. pictus胚胎LpMT 1启动子是一个强大的，可诱导的，混杂的启动子用于驱动海胆胚胎中异源基因的表达。
The 5'-flanking region of the metallothionein (MT) gene LpMT1 of the sea urchin Lytechinus pictus includes three copies of a conserved sequence that includes the metal-responsive element (MRE) consensus core sequence required for heavy metal induction of other MT genes, a GC box, a G box of a putative basal level enhancer element which includes another MRE core element, and a poly(C) tract. A fragment of LpMT1 DNA from nucleotides + 31 to - 309 fused to a chloramphenicol acetyltransferase reporter gene was inducible with cadmium after injecton into L. pictus embryos. This induced activity was greatly reduced in a deletion mutant which retained only 195 base pairs of 5'-flanking sequence, including the proximal pair of MREs and the G box, but excluding the poly(C) tract, GC box, and distal MRE. A potent human hMT-II(A) gene promoter is marginally functional in L. pictus embryos. In contrast, the LpMT1 promoter is active in HeLa cells and in embryos of the sea urchin Strongylocentrotus purpuratus. The hMT-II(A) gene may lack a cis-acting sequence clement required for expression of MT genes in L. pictus embryos. The LpMT1 promoter is a powerful, inducible, promiscuous promoter useful for driving the expression of heterologous genes in sea urchin embryos.