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.
Artigo
em Chinês
| WPRIM
| ID: wpr-256169
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:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Proteínas de Bactérias
/
Proteínas Recombinantes
/
DNA
/
Cromatografia de Afinidade
/
Adenosina Trifosfatases
/
Pareamento Incorreto de Bases
/
Proteínas de Escherichia coli
/
Proteínas de Ligação a DNA
/
Reparo do DNA
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2002
Tipo de documento:
Artigo
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