Authors: Narasimha Anaganti Bhakti Basu Shree Kumar Apte
Publish Date: 2016/05/05
Volume: 41, Issue: 2, Pages: 193-203
Abstract
A third generation promoter probe shuttle vector pKG was constructed using the green fluorescent protein as a reporter for in situ evaluation of Deinococcal promoter activity in Escherichia coli or Deinococcus radiodurans The construct yielded zero background fluorescence in both the organisms in the absence of promoter sequences Fifteen Deinococcal promoters either harbouring Radiation and Desiccation Response Motif RDRM or not were cloned in vector pKG Only the RDRMpromoter constructs displayed i gamma radiation inducible GFP expression in D radiodurans following gamma irradiation ii DdrOmediated repression of GFP expression in heterologous E coli or iii abolition in GFP induction following gamma irradiation in pprI mutant of D radiodurans Utility of pKG vector for realtime in situ assessment of Deinococcal promoter function was thus successfully demonstratedThe authors wish to thank KW Minton and MJ Daly Uniformed Services University of the Health Sciences Bethesda MD for providing Deinococcus radiodurans strain R1 ATCC BAA816 ME Lidstrom Departments of Chemical Engineering and Microbiology University of Washington Seattle for providing plasmid pRAD1 and JW Golden Section of Molecular Biology UC San Diego USA for the gift of vector pAM1956 SKA thanks the Department of Science and Technology India for the award of Sir JC Bose National Fellowship SERB/F/2569/201314 dated 29072013 and Department of Atomic Energy India for the award of Raja Ramanna Fellowship DAE/10/120/2014/RRFRDII/3208 dated 11032015
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