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Expression, Purification, Crystallization and X-Ray Crystallographic Analysis of MoDabb1 from Magnaporthe oryzae

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Abstract

Protein acetylation is one of the most common post-translational modifications. Many acetylated proteins in Magnaporthe oryzae play key roles in vegetative growth and pathogenicity. MoDabb1 from M. oryzae was also identified to be an acetylated protein, containing a Dabb domain with unknown function. To elucidate the function of this protein and the effect of acetylation on this protein, a native and selenomethionine-substituted MoDabb1 were expressed in Escherichia coli and purified to homogeneity. Crystals were obtained using sitting-drop vapour-diffusion method. Crystals of native and selenomethionine-substituted protein were diffracted to a resolution of 1.74 and 1.98 Å and both belonged to the sp. gr. P42212. Matthews coefficient analysis indicated two molecules in an asymmetric unit with a Vm value of 2.41 and a corresponding solvent content of 49.03%.

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ACKNOWLEDGMENTS

We are grateful to Dr. Arie Geerlof at EMBO for kindly supplying the expression vector pHAT2. We also thank all the staffs from BL18U1 and BL19U1 beamline of National Facility for Protein Science Shanghai (NFPS) at Shanghai Synchrotron Radiation Facility (SSRF) for help with crystal screening and data collection. This research was supported by the National Key Basic Research and Development Program (no. 2017YFD0201705), the National Natural Science Foundation of China (NSFC) (nos. 31471735 and 31772112), and the Taishan Scholar Construction Foundation of Shandong Province (no. 6631114314).

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Authors and Affiliations

  1. College of Plant Health and Medicine, Key Lab of Integrated Crop Disease and Pest Management of Shandong Province, Qingdao Agricultural University, 266109, Qingdao, Shandong, China

    Y. Zhao, M. Chi, W. Liang & J. Huang

  2. College of Plant Protection, MOA Key Laboratory of Plant Pathology, China Agricultural University, 100193, Haidian District, Beijing, China

    Y. Zhao, X. Zhang, S. Wang & J. Liu

Authors
  1. Y. Zhao
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  2. M. Chi
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  3. X. Zhang
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  4. S. Wang
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  5. J. Liu
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  6. W. Liang
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  7. J. Huang
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Correspondence to J. Huang.

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Zhao, Y., Chi, M., Zhang, X. et al. Expression, Purification, Crystallization and X-Ray Crystallographic Analysis of MoDabb1 from Magnaporthe oryzae. Crystallogr. Rep. 64, 1112–1116 (2019). https://doi.org/10.1134/S1063774519070307

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  • DOI: https://doi.org/10.1134/S1063774519070307

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