两种人类芳基硫酸转移酶的细菌表达。

Bacterial expression of two human aryl sulfotransferases.

作者信息

Bidwell L M, Gillam E M, Gaedigk A, Zhu X, Grant D, McManus M E

机构信息

Department of Physiology and Pharmacology, The University of Queensland, Brisbane, Australia.

出版信息

Chem Biol Interact. 1998 Feb 20;109(1-3):137-41. doi: 10.1016/s0009-2797(97)00128-2.

DOI:10.1016/s0009-2797(97)00128-2

PMID:9566741

Abstract

The effect of replacing a single codon in the N-terminal of human aryl sulfotransferase (HAST) 1 and 3 with one that is more commonly found in E. coli genes was assessed. The pKK233-2 E. coli expression vector was employed and the polymerase chain reaction (PCR) was used to introduce the 5' nucleotide substitution, at the same time maintaining the fidelity of the amino acid sequence. The data indicates that this change had a minimal effect on protein production, subcellular localization or, in the case of HAST3, catalytic activity. In general, the pKK233-2 E. coli vector has been less than optimal for expressing human sulfotransferase cDNAs.

摘要

评估了将人芳基磺基转移酶（HAST）1和3 N端的单个密码子替换为大肠杆菌基因中更常见密码子的效果。使用了pKK233-2大肠杆菌表达载体，并通过聚合酶链反应（PCR）引入5'核苷酸取代，同时保持氨基酸序列的保真度。数据表明，这种变化对蛋白质产生、亚细胞定位或（就HAST3而言）催化活性的影响极小。一般来说，pKK233-2大肠杆菌载体在表达人磺基转移酶cDNA方面一直不太理想。