[Cloning and expression of spinach glycolate oxidase in Escherichia coli].
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.
Jin
Jian-Feng
JF
Department of Biochemical Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Tan
Tian-Wei
TW
Su
Guo-Fu
GF
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli; D015870:Gene Expression / Q000187:drug effects; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D018724:Spinacia oleracea / Q000201:enzymology* / Q000235:genetics
D007544:Isopropyl Thiogalactoside; D000429:Alcohol Oxidoreductases; C020821:glycollate oxidase