Cloning and expression of spinach glycolate oxidase in Escherichia coli.
- Author:
Jian-Feng JIN
1
;
Tian-Wei TAN
;
Guo-Fu SU
Author Information
1. Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
- Publication Type:Journal Article
- MeSH:
Alcohol Oxidoreductases;
genetics;
isolation & purification;
metabolism;
Cloning, Molecular;
Escherichia coli;
Gene Expression;
drug effects;
Isopropyl Thiogalactoside;
pharmacology;
Spinacia oleracea;
enzymology;
genetics
- From:Chinese Journal of Biotechnology
2002;18(2):212-215
- CountryChina
- Language:Chinese
-
Abstract:
The cDNA coding spinach glycolate oxidase (GO) was amplified by RT-PCR using the total RNA of spinach leaves as the template, and was cloned into cloning vector pMD18-T. After the DNA sequence was determined, the go gene was subcloned into E. coli expression vector pBV220, pET-22b(+), pTIG-Trx and pThioHisC. SDS-PAGE analysis revealed that the recombinant E. coli BL21 (DE3) (pTIG-Trx-GO) and E. coli BL21 (DE3) (pET-22b(+)-GO) expressed the predicted 38 kD glycolate oxidase, and the enzyme activity was also detected.
