Expression of human glutathione S-transferase A1, P1 and T1 in Escherichia coli / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 168-174, 2014.
Article
Dans Chinois
| WPRIM
| ID: wpr-336723
ABSTRACT
<p><b>OBJECTIVE</b>To construct the vectors of human glutathione S-transferase A1 (GSTA1), P1 (GSTP1), T1(GSTT1) genes and express in Escherichia coli (E. coli).</p><p><b>METHODS</b>Human GSTA1, GSTP1 and GSTT1 gene whole length cDNAs were amplified by RT-PCR and then subcloned into pET-28a(+) vectors. The proteins were expressed in E. coli BL21(DE3). After purified by Ni2+ affinity chromatography, the enzymatic activities of GSTs were measured with 1-chloro-2,4 -dinitrobenzene (CDNB) as substrate.</p><p><b>RESULTS</b>The correct GSTA1, GSTP1 and GSTT1 genes were cloned. And soluble GSTA1, GSTT1, GSTP1 proteins were expressed in E.coli. After purification, GSTA1, GSTT1 and GSTP1 showed good enzymatic activities, which were 17.55, 0.02, 18.75 μmol·min-1·mg-1, respectively.</p><p><b>CONCLUSION</b>The expression plasmids for GSTA1, GSTT1 and GSTP1 have been constructed and the recombinant proteins are expressed successfully.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Protéines recombinantes
/
ADN complémentaire
/
RT-PCR
/
Escherichia coli
/
Glutathione S-transferase pi
/
Vecteurs génétiques
/
Génétique
/
Glutathione transferase
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Zhejiang University. Medical sciences
Année:
2014
Type:
Article
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