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可在哺乳动物细胞中表达的人类末端脱氧核苷酸转移酶cDNA分析

Analysis of human terminal deoxynucleotidyl transferase cDNA expressible in mammalian cells.

作者信息

Koiwai O, Kaneda T, Morishita R

出版信息

Biochem Biophys Res Commun. 1987 Apr 14;144(1):185-90. doi: 10.1016/s0006-291x(87)80493-x.

Abstract

Human Molt3 cDNA library was constructed using pcD vector system which permits the expression of cDNA inserts in mammalian cells. Nearly full-length human terminal deoxynucleotidyltransferase (TdT) cDNA was cloned using a fragment of bovine TdT cDNA as a probe. The human TdT cDNA contains an open reading frame of 1,557 bp coding for 519 amino acids, including 31 bp and 341 bp from 5' and 3' untranslated regions, respectively. The TdT cDNA was transfected into COS7 monkey fibroblasts directed the synthesis of enzymatically active protein of Mr 59,495. The cloned TdT cDNA hybridized with poly A+ RNAs of 2,100 b and 3,300 b from stable T-cell leukemia Molt3 and Molt4 cells.

摘要

使用允许cDNA插入片段在哺乳动物细胞中表达的pcD载体系统构建了人Molt3 cDNA文库。以牛TdT cDNA片段为探针克隆了几乎全长的人末端脱氧核苷酸转移酶(TdT)cDNA。人TdT cDNA包含一个1557 bp的开放阅读框,编码519个氨基酸,分别包括来自5'和3'非翻译区的31 bp和341 bp。将TdT cDNA转染到COS7猴成纤维细胞中,指导合成Mr 59,495的酶活性蛋白。克隆的TdT cDNA与稳定的T细胞白血病Molt3和Molt4细胞中2100 b和3300 b的聚A + RNA杂交。

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