Characterisation and expression of the carbamate kinase gene from Giardia intestinalis

被引：14

作者：

Minotto, L ^{[1
]}

Tutticci, EA ^{[1
]}

Bagnara, AS ^{[1
]}

Schofield, PJ ^{[1
]}

Edwards, MR ^{[1
]}

机构：

[1] Univ New S Wales, Sch Biol & Mol Genet, Sydney, NSW 2052, Australia

来源：

MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 1999年 / 98卷 / 01期

基金：

澳大利亚研究理事会; 英国医学研究理事会;

关键词：

carbamate kinase; gene cloning and sequencing; gene expression; characterisation of recombinant carbamate kinase; Giardia intestinalis;

D O I：

10.1016/S0166-6851(98)00141-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The arginine dihydrolase pathway in Giardia intestinalis produces energy via the carbamate kinase (CBK, ATP:carbamate phosphotransferase, EC 2.7.2.2) reaction. Characterisation of the CBK gene from the Portland 1 strain indicated that it is located on either chromosome 3 or 4, does not appear to contain introns and is expressed in both the trophozoite and early cyst stages. Heterologous expression of CBK in Escherichia cell, using the pQE-30 expression system (QIAGEN), enabled a one-step purification of the recombinant enzyme via affinity chromatography, The expressed protein was identified by enzyme assay and mass spectrometry. The native and recombinant forms of the enzyme have similar physical properties and the recombinant enzyme appears to be active as the homodimer. (C) 1999 Published by Elsevier Science B.V. All rights reserved.

引用

页码：43 / 51

页数：9

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