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一种42千道尔顿大鼠肾胆碱/乙醇胺激酶的互补DNA序列。

Complementary DNA sequence for a 42 kDa rat kidney choline/ethanolamine kinase.

作者信息

Aoyama C, Nakashima K, Matsui M, Ishidate K

机构信息

Department of Molecular Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, Chiyodaku, Tokyo 101, Japan.

出版信息

Biochim Biophys Acta. 1998 Feb 5;1390(1):1-7. doi: 10.1016/s0005-2760(97)00177-x.

Abstract

By means of peptide sequence information, several cDNA clones encoding a 42 kDa choline/ethanolamine kinase were isolated from a rat kidney cDNA library. Eight clones were sequenced with all of them resulting in identical overlapping nucleotide sequences. Four of them possessed entire open reading frame which could encode 394 amino acids with a calculated molecular size of 45 100. The predicted amino acid sequence contained all of the internal peptide fragment sequences derived from the purified 42 kDa enzyme. When the open reading frame was introduced into pGEX-2T vector and transfected into E. coli cells, a significant choline/ethanolamine kinase activity did appear in the cell lysate. A homology comparison with the previously reported choline kinase cDNAs (CKR1 and CKR2) from rat liver showed 66%-68% in entire nucleotide sequences and 57%-59% in amino acid sequences, indicating that the cloned cDNA here must be a novel CK/EK gene product. (c) 1998 Elsevier Science B. V.

摘要

通过肽序列信息,从大鼠肾脏cDNA文库中分离出几个编码42 kDa胆碱/乙醇胺激酶的cDNA克隆。对8个克隆进行了测序,所有克隆都产生了相同的重叠核苷酸序列。其中4个克隆拥有完整的开放阅读框,可编码394个氨基酸,计算出的分子大小为45100。预测的氨基酸序列包含了从纯化的42 kDa酶衍生而来的所有内部肽片段序列。当将开放阅读框导入pGEX - 2T载体并转染到大肠杆菌细胞中时,细胞裂解物中确实出现了显著的胆碱/乙醇胺激酶活性。与先前报道的大鼠肝脏胆碱激酶cDNA(CKR1和CKR2)进行同源性比较,发现整个核苷酸序列的同源性为66% - 68%,氨基酸序列的同源性为57% - 59%,这表明此处克隆的cDNA必定是一种新的CK/EK基因产物。(c)1998爱思唯尔科学出版社B.V.

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