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KMID : 0880220070450020175
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Journal of Microbiology
2007 Volume.45 No. 2 p.175 ~ p.178
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Recombinant Expression and Purification of Functional XorII, a Restriction Endonuclease from Xanthomonas oryzae pv. oryzae
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Hwang Dong-Kyu
Cho Jae-Yong
Chae Young-Kee
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Abstract
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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°C 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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KEYWORD
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XorII, restriction endonuclease, Xanthomonas oryzae pv. oryzae
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