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Molecular characterization of DNA repair protein Ku70 from Vitis vinifera and its purification from transgenic tobacco

Abstract

The DNA double strand break repair in plants is preferentially by non homologous end joining (NHEJ) pathway. A key protein of NHEJ pathway is Ku70. We have identified Ku70 homolog (VvKu70) from grapevine genome database. In this report we characterize a Ku70 homologue from Vitis vinifera cv. Mango. The VvKu70 expression was found to increase strongly in response to gamma radiation. The transcript level of VvKu70 was found to increase up to 36 h in gamma irradiated shoots of grapevine. The expression of VvKu70 was found in many organs like stem, leaves and roots. A GFP fused VvKu70 protein was found to be nuclear localized which indicates that the VvKu70 is a nuclear localized protein. The VvKu70 identified by in silico approaches is present as a single copy number in V. vinifera cv. Mango genome. The VvKu70-GFP fused protein possesses ATPase activity and fails to bind dsDNA but binds ssDNA.

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Acknowledgments

Authors thank Dr. S.F. D’Souza, Head, Nuclear Agriculture and Biotechnology Division, BARC for his constant encouragement. Authors would also like to thank Mr. Upendra Singh Shekhawat, Dr. T.R. Ganapathi and Dr. S.B. Ghosh for scientific inputs.

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Correspondence to Himanshu Tak.

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Tak, H., Mhatre, M. Molecular characterization of DNA repair protein Ku70 from Vitis vinifera and its purification from transgenic tobacco. Transgenic Res 22, 839–848 (2013). https://doi.org/10.1007/s11248-013-9690-y

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Keywords

  • Vitis spp.
  • Ku70
  • Malachite green
  • GFP
  • DNA repair