High expression and identification of DNA mismatch repair gene mutS in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 536-540, 2002.
Article
en Zh
| WPRIM
| ID: wpr-256169
Biblioteca responsable:
WPRO
ABSTRACT
DNA mismatch repair gene mutS (2.56 kb) was PCR modified and cloned into a secretive prokaryotic expression vector pET32a (+) which carries a N-terminal His.tag + and thioredoxin sequence. MutS protein was expressed with high level after IPTG induction using the strain E. coli AD494(DE3). SDS-PAGE revealed that the expected protein with a molecular weight of 108 kD which is about 35% of the total bacterial proteins is almost soluble. The expected protein was purified directly by immobilized metal (Ni2+) chelation affinity chromatography and the purity is over 90%. MutS protein activity verified using mismatch DNA showed that the expression product can recognize and bind to base-pair mismatch specifically.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Farmacología
/
Proteínas Bacterianas
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Proteínas Recombinantes
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ADN
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Cromatografía de Afinidad
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Adenosina Trifosfatasas
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Disparidad de Par Base
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Proteínas de Escherichia coli
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Proteínas de Unión al ADN
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Reparación del ADN
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2002
Tipo del documento:
Article