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      https://www.ias.ac.in/article/fulltext/jbsc/041/02/0193-0203

    • Keywords

       

      D. radiodurans; GFP fluorescence; promoter probe shuttle vector; radiation-responsive gene expression

    • 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 RDRM-promoter constructs displayed (i) gamma radiation inducible GFP expression in D. radiodurans, following gamma irradiation, (ii) DdrO-mediated 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 real-time in situ assessment of deinococcal promoter function was, thus, successfully demonstrated.

    • Author Affiliations

       

      Narasimha Anaganti1 Bhakti Basu1 Shree Kumar Apte1

      1. Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India
    • Dates

       
    • Supplementary Material

       
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