Human thymidylate synthase gene: isolation of phage clones which cover a functionally active gene and structural analysis of the region upstream from the translation initiation codon.
Human thymidylate synthase gene: isolation of phage clones which cover a functionally active gene and structural analysis of the region upstream from the translation initiation codon.
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人胸苷酸合酶基因：分离包含功能活性基因的噬菌体克隆，并对翻译起始密码子上游区域进行结构分析。
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
Takeshi Seno
中科院分区：
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作者：
K. Takeishi;S. Kaneda;Dai Ayusawa;K. Shimizu;Osamu Gotoh;Takeshi Seno
两个部分编码人胸苷酸合酶（TS）的基因组DNA片段[EC 2.1.1.45]先前已在来自小鼠细胞的λ噬菌体中克隆，但对小鼠TS阴性突变细胞没有转化活性。在本研究中，克隆了人TS的另外的基因组DNA，并证明其与先前克隆的两种DNA之一组合具有转化活性，并产生人TS mRNA。两个转化基因组DNA重叠，覆盖了总共23 kb的区域。使用来自这些DNA之一的片段，分析了TS基因的ATG起始密码子周围1.2 kb区域的结构与该基因的调控序列的关系。序列测定表明存在一个不寻常的反向重复序列，由一个28 bp的序列和一个相同长度的反向序列的三重串联重复组成。这些序列可以形成三种可能的、稳定的茎环结构，它们可以相互转化。基于S1核酸酶图谱数据和一系列间接证据，我们推断出反向序列中的两个主要mRNA帽位点。人和小鼠序列上游的ATG起始密码子的比较揭示了许多显着的块的序列同源性，特别是在推导的帽位点周围的区域。
Two genomic DNA fragments partially encoding human thymidylate synthase (TS) [EC 2.1.1.45] were previously cloned in lambda phage from the mouse cell transformant, but had no transforming activity on mouse TS-negative mutant cells. In this study, an additional genomic DNA for human TS was cloned and demonstrated to have the transforming activity in combination with one of the two previously cloned DNAs and to produce human TS mRNA. The two transforming genomic DNAs overlapped and covered a region of 23 kb in total. Using fragments from one of these DNAs, the structure of the 1.2-kb region around the ATG initiator codon of the TS gene was analyzed in relation to regulatory sequences of the gene. Sequence determination demonstrated the presence of an unusual inverted repeat consisting of a triple tandem repeat of a 28-bp sequence and an inverted sequence of the same length. These sequences can form three possible, stable, stem-loop structures, which may be interconvertible. Based on S1 nuclease mapping data and a line of circumstantial evidence, we deduced two major mRNA cap sites within the inverted sequence. Comparison of the human and mouse sequences upstream from the ATG initiator codon revealed many significant blocks of sequence homology, especially in the regions around the deduced cap sites.