| Title | Recombinant Expression and Purification of Functional XorII, a Restriction Endonuclease from Xanthomonas oryzae pv. oryzae |
|---|---|
| Author | Dong Kyu Hwang1, Jae-Yong Cho2, and Young Kee Chae1* |
| Address | 1Department of Chemistry, Sejong University, Seoul 143-747, Republic of Korea, 2Department of Bioindustry and Technology, Sangji University, Gangwon-Do 220-702, Republic of Korea |
| Bibliography | Journal of Microbiology, 45(2),175-178, 2007, |
| DOI | |
| Key Words | XorII, restriction endonuclease, Xanthomonas oryzae pv. oryzae |
| 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°C in 50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, and 1 mM dithiothreitol at a pH of 7.9. |
| Download PDF | 45(2)-14.pdf |