Molecular cloning and functional analysis of the Arabidopsis thaliana DNA ligase I homologue.
Molecular cloning and functional analysis of the Arabidopsis thaliana DNA ligase I homologue.
复制标题
拟南芥 DNA 连接酶 I 同源物的分子克隆和功能分析。
DOI：
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
C. M. Bray
中科院分区：
文献类型：
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作者：
Richard M. Taylor;M. J. Hamer;J. Rosamond;C. M. Bray
已使用简并PCR方法从拟南芥中分离出编码DNA连接酶I同源物的cDNA。该cDNA的ORF编码790个残基的氨基酸序列，代表理论分子量为87.8 kDa且等电点（pi）为8.20的蛋白质。A的对齐。拟南芥DNA连接酶蛋白序列与来自人（Homo sapiens）、鼠（Mus musculus）、爪蟾（Xenopus laevis）和酵母裂殖酵母（Schizoschiomycespombe）和酿酒酵母（Saccharomycescerevisiae）的DNA连接酶序列显示出良好的序列同源性（42-45%同一性，61-66%相似性），特别是在活性位点周围。序列数据表明拟南芥DNA连接酶是动物DNA连接酶I物种的同源物。cDNA克隆的功能分析表明其能够补充S.酿酒酵母cdc 9突变体DNA连接酶活性缺陷，证实克隆的序列编码活性DNA连接酶。DNA连接酶转录水平在A.拟南芥幼苗对一段时间增强UV-B辐射诱导的DNA损伤的反应。然而，DNA连接酶mRNA转录物的细胞水平确实与植物细胞的复制状态相关。
A cDNA encoding the DNA ligase I homologue has been isolated from Arabidopsis thaliana using a degenerate PCR approach. The ORF of this cDNA encodes an amino acid sequence of 790 residues, representing a protein with a theoretical molecular mass of 87.8 kDa and an isoelectric point (pi) of 8.20. Alignment of the A. thaliana DNA ligase protein sequence with the sequence of DNA ligases from human (Homo sapiens), murine (Mus musculus), clawed toad (Xenopus laevis) and the yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae showed good sequence homology (42-45% identity, 61-66% similarity), particularly around the active site. Sequence data indicate that the Arabidopsis DNA ligase is the homologue of the animal DNA ligase I species. Functional analysis of the cDNA clone demonstrated its ability to complement the conditional lethal phenotype of an S. cerevisiae cdc9 mutant defective in DNA ligase activity, confirming that the cloned sequence encodes an active DNA ligase. The level of the DNA ligase transcript was not increased in A. thaliana seedlings in response to DNA damage induced by a period of enhanced UV-B irradiation. However, the cellular level of the DNA ligase mRNA transcript does correlate with the replicative state of plant cells.
酿酒酵母有丝分裂提取物中两种不同的 DNA 连接酶活性。
影响因子：
14.9
作者：
Ramos,W;Tappe,N;Talamantez,J;Friedberg,EC;Tomkinson,AE
通讯作者：
Tomkinson,AE