一种编码酚磺基转移酶的牛基因的分离与鉴定。

Isolation and characterization of a bovine gene encoding phenol sulfotransferase.

作者信息

Henry T, Kliewer B, Palmatier R, Ulphani J S, Beckmann J D

机构信息

Department of Internal Medicine, University of Nebraska Medical Center, Omaha 68198-5300, USA.

出版信息

Gene. 1996 Oct 3;174(2):221-4. doi: 10.1016/0378-1119(96)00083-2.

DOI:10.1016/0378-1119(96)00083-2

PMID:8890738

Abstract

Phenol sulfotransferases (PST) esterify hydroxylated aromatic compounds with sulfate, and therefore play a role in the metabolism of xenobiotics. In this investigation, a bovine PST gene of 2372 bp was amplified from genomic DNA. Sequence overlap with the cognate cDNA revealed seven exons, with all introns containing GT/AG splicing donor/acceptor site. This product was subcloned into an SV40-based expression vector and transfected into WI-26 human lung fibroblasts. Immunoblot analyses revealed production of the anticipated 32-kDa protein, and the active enzyme displayed steady-state kinetic properties consistent with the enzyme characterized in bovine lung (apparent Km = 6.5 microM for 2-naphthol).

摘要

酚磺基转移酶（PST）可使羟基化芳香族化合物与硫酸盐发生酯化反应，因此在异生素代谢中发挥作用。在本研究中，从基因组DNA中扩增出一个2372 bp的牛PST基因。与同源cDNA的序列重叠显示有7个外显子，所有内含子均含有GT/AG剪接供体/受体位点。该产物被亚克隆到基于SV40的表达载体中，并转染到WI-26人肺成纤维细胞中。免疫印迹分析显示产生了预期的32 kDa蛋白质，并且活性酶表现出与牛肺中鉴定的酶一致的稳态动力学特性（对2-萘酚的表观Km = 6.5 microM）。