编码小鼠DNA连接酶I的cDNA的克隆与序列分析。

Cloning and sequence analysis of a cDNA coding for the murine DNA ligase I enzyme.

作者信息

Savini E, Biamonti G, Ciarrocchi G, Montecucco A

机构信息

Istituto di Genetica Biochimica ed Evoluzionistica, C.N.R., Pavia, Italy.

出版信息

Gene. 1994 Jul 8;144(2):253-7. doi: 10.1016/0378-1119(94)90386-7.

DOI:10.1016/0378-1119(94)90386-7

PMID:8039710

Abstract

A complementary DNA (2961 bp) containing the complete coding sequence for murine DNA ligase I was isolated from a mouse fibroblast cDNA library using a cDNA encoding the human protein as a probe. An open reading frame of 2748 bp, encoding a protein of 916 amino acids (aa), was identified. Northern blot analysis of total RNA extracted from mouse fibroblasts showed a single band with a mobility corresponding to a size of 3.2 kb whose level increases upon serum stimulation of quiescent mouse NIH-3T3 cells. Alignment of the murine and human deduced aa sequences showed an overall 83% identity, that rises to 91% if only the sequence on the C-terminal portion of the protein containing the active site is considered.

摘要

使用编码人类蛋白质的互补DNA作为探针，从小鼠成纤维细胞cDNA文库中分离出一个包含小鼠DNA连接酶I完整编码序列的互补DNA（2961碱基对）。鉴定出一个2748碱基对的开放阅读框，其编码一个由916个氨基酸组成的蛋白质。对从小鼠成纤维细胞中提取的总RNA进行Northern印迹分析，结果显示有一条迁移率对应于3.2 kb大小的单带，在血清刺激静止的小鼠NIH-3T3细胞后，其水平升高。小鼠和人类推导的氨基酸序列比对显示，总体一致性为83%，如果仅考虑蛋白质C端部分包含活性位点的序列，一致性则升至91%。