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人类β-1,4-半乳糖基转移酶cDNA克隆序列分析

Analysis of the sequences of human beta-1,4-galactosyltransferase cDNA clones.

作者信息

Chatterjee S K, Mukerjee S, Tripathi P K

机构信息

Department of Obstetrics and Gynecology, University of Kentucky, Lexington 40536-0096, USA.

出版信息

Int J Biochem Cell Biol. 1995 Mar;27(3):329-36. doi: 10.1016/1357-2725(94)00062-g.

Abstract

UDP-galactose:beta-1,4 N-acetyl glucosamine galactosyltransferase (4 beta GT) is a promising tumor marker for ovarian cancer. To study the role of 4 beta GT in malignant transformation at the molecular level human 4 beta GT cDNA and genomic clones were isolated and analyzed. For the isolation of 4 beta GT cDNA and genomic clones, a human fetal liver cDNA library in lambda gt11 and a human genomic library in EMBL-3B vectors respectively were screened using a 4 beta GT cDNA insert as the probe. Complete sequence of the cDNA clones were determined by subcloning in plasmid vectors, and compared with the published sequence of human liver 4 beta GT. Presence of various 4 beta GT exons in the genomic clones were determined by Southern blot analysis using specific oligodeoxynucleotide probes. Among the 5 cDNA clones isolated, 2 clones GTN 6 and GTN 17 were sibling clones and had a nucleotide sequence identical to the published 4 beta GT cDNA sequence, except at the 3'-end, where these clones had 7 unique nucleotide sequences. One cDNA clone, GTN2 also had a nucleotide sequence identical to that of 4 beta GT, except for 3 G residues at the 5'-end. One cDNA clone, GTN 1, had a unique sequence at the 5'-end comprising of 74 nucleotides. Another clone, GTN 20, was unrelated to 4 beta GT. Analysis of genomic clones showed that 4 beta GT exons 3, 4, 5 and 6 were present in a 14 kb genomic clone, EMGT-4. Exon 1 was present in a separate 16 kb clone, EMGT-6.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

UDP-半乳糖:β-1,4-N-乙酰葡糖胺半乳糖基转移酶(4βGT)是一种很有前景的卵巢癌肿瘤标志物。为了在分子水平研究4βGT在恶性转化中的作用,分离并分析了人4βGT cDNA和基因组克隆。为了分离4βGT cDNA和基因组克隆,分别使用4βGT cDNA插入片段作为探针,筛选λgt11载体中的人胎肝cDNA文库和EMBL-3B载体中的人基因组文库。通过亚克隆到质粒载体中确定cDNA克隆的完整序列,并与已发表的人肝4βGT序列进行比较。使用特异性寡脱氧核苷酸探针通过Southern印迹分析确定基因组克隆中各种4βGT外显子的存在。在分离出的5个cDNA克隆中,2个克隆GTN 6和GTN 17是同胞克隆,其核苷酸序列与已发表的4βGT cDNA序列相同,但在3'-末端除外,这些克隆在该末端有7个独特的核苷酸序列。一个cDNA克隆GTN2的核苷酸序列也与4βGT相同,只是在5'-末端有3个G残基除外。一个cDNA克隆GTN 1在5'-末端有一个由74个核苷酸组成的独特序列。另一个克隆GTN 20与4βGT无关。基因组克隆分析表明,4βGT外显子3、4、5和6存在于一个14 kb的基因组克隆EMGT-中。外显子1存在于一个单独的16 kb克隆EMGT-6中。(摘要截短至250字)

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