Abstract
Almost complete sequence specific 1H, 13C and 15N resonance assignments of a mutant of UV inducible transcript (S55A-UVI31+) from Chlamydomonas reinhardtii are reported, as a prelude to its structural and functional characterization. Site-directed mutagenesis of uvi31+ was carried out using complementary mutation harbouring oligonucleotides for S55A mutant. The resulting mutant was sequenced and then S55A mutant of uvi31+ cDNA were expressed in E. coli, and purification were carried out from where the protein was purified to high homogeneity. The point mutation S55A in UVI31+ results in a significant enhancement in its DNA endonuclease activity as compared to its wild type protein.
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Acknowledgments
The facilities provided by National Facility for High Field NMR at Tata Institute of Fundamental Research, Mumbai, supported by Department of Science and Technology, New Delhi, Department of Biotechnology, New Delhi, Council of Scientific and Industrial Research, New Delhi and Tata Institute of Fundamental Research, Mumbai are acknowledged. KVRC and BJ thank DST for their JC Bose Fellowships.
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Singh, H., Rao, B.J. & Chary, K.V.R. 1H, 13C and 15N NMR assignments of a mutant of UV inducible transcript (S55A-UVI31+) from Chlamydomonas reinhardtii . Biomol NMR Assign 8, 371–374 (2014). https://doi.org/10.1007/s12104-013-9520-4
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DOI: https://doi.org/10.1007/s12104-013-9520-4