Purification of Recombinant N-terminal Histidine-Tagged Arabidopsis thaliana Phosphoglycolate Phosphatase 1, Glycolate Oxidase 1 and 2, and Hydroxypyruvate Reductase 1.
Methods Mol Biol
; 2792: 97-111, 2024.
Article
em En
| MEDLINE
| ID: mdl-38861081
ABSTRACT
To measure the kinetic properties of photorespiratory enzymes, it is necessary to work with purified proteins. Protocols to purify photorespiratory enzymes from leaves of various plant species require several time-consuming steps. It is now possible to produce large quantities of recombinant proteins in bacterial cells. They can be rapidly purified as histidine-tagged recombinant proteins by immobilized metal affinity chromatography using Ni2+-NTA-agarose. This chapter describes protocols to purify several Arabidopsis thaliana His-tagged recombinant photorespiratory enzymes (phosphoglycolate phosphatase, glycolate oxidase, and hydroxypyruvate reductase) from Escherichia coli cell cultures using two bacterial strain-plasmid systems BL21(DE3)-pET and LMG194-pBAD.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
Monoéster Fosfórico Hidrolases
/
Proteínas de Arabidopsis
/
Oxirredutases do Álcool
/
Escherichia coli
/
Hidroxipiruvato Redutase
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article