Recombinant expression and purification of functional XorII, a restriction endonuclease from Xanthomonas oryzae pv. oryzae.
J Microbiol
; 45(2): 175-8, 2007 Apr.
Article
en En
| MEDLINE
| ID: mdl-17483805
ABSTRACT
An endonuclease from Xanthomonas oryzae pathovar oryzae KACC 10331, XorII, was recombinantly produced in Escherichia coli using a T7 system. XorII was purified using a combination of ion exchange and immobilized metal affinity chromatography (IMAC). An optimized washing protocol was carried out on an IMAC in order to obtain a high purity product. The final amount of purified XorII was approximately 2.5 mg/L of LB medium. The purified recombinant XorII was functional and showed the same cleavage pattern as PvuI. The enzyme activity tested the highest at 25 degrees in 50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, and 1 mM dithiothreitol at a pH of 7.9.
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Banco de datos:
MEDLINE
Asunto principal:
Proteínas Bacterianas
/
Xanthomonas
/
Enzimas de Restricción del ADN
Idioma:
En
Año:
2007
Tipo del documento:
Article