A 12-base-pair DNA motif that is repeated several times in metallothionein gene promoters confers metal regulation to a heterologous gene.
A 12-base-pair DNA motif that is repeated several times in metallothionein gene promoters confers metal regulation to a heterologous gene.
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DOI：
10.1073/pnas.81.23.7318
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发表时间：
1984-12
影响因子：
11.1
通讯作者：
G. Stuart;P. Searle;Howard Y. CHENt;R. L. BRINSTERt;R. Palmiter
中科院分区：
文献类型：
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作者：
G. Stuart;P. Searle;Howard Y. CHENt;R. L. BRINSTERt;R. Palmiter
为了定义参与小鼠金属硫蛋白-I (MT-I) 基因启动子功能和金属调控的 DNA 序列，我们将 MT-I 基因的 5' 侧翼序列与病毒胸苷激酶 (TK) 基因的编码序列融合。构建了一系列 MT-1 启动子的 5' 缺失、3' 缺失、接头扫描和内部缺失突变体，并通过显微注射到小鼠卵中进行分析。结果表明至少两个相关的启动子元件可以独立地赋予一些金属调节作用。这些对调节影响最严重的突变会影响 12 碱基对保守序列，该序列在小鼠 MT-I 和其他 MT 启动子中重复多次。为了测试该序列的调节功能，将其合成为一对互补寡核苷酸并插入到 TK 基因的启动子中。该序列的单个插入赋予TK启动子有限的金属调节，而具有两个单独插入的构建体与MT-I启动子一样有效地受到调节。
To define DNA sequences involved in mouse metallothionein-I (MT-I) gene promoter function and metal regulation, we fused the 5' flanking sequences of the MT-I gene to the coding sequences of a viral thymidine kinase (TK) gene. A series of 5' deletion, 3' deletion, linker-scanning, and internal deletion mutants of the MT-I promoter was constructed and assayed by microinjection into mouse eggs. The results indicate that at least two related promoter elements can confer some metal regulation independently. Those mutations that had the most severe effect on regulation impinge on a 12-base-pair conserved sequence that is repeated several times within the mouse MT-I and other MT promoters. To test the regulatory function of this sequence, it was synthesized as a pair of complementary oligonucleotides and inserted into the promoter of the TK gene. A single insertion of this sequence conferred limited metal regulation onto the TK promoter, whereas a construct with two separate inserts was regulated as efficiently as the MT-I promoter.