title: In situ real-time evaluation of radiation-responsive promoters in the extremely radioresistant microbe Deinococcus radiodurans.
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.
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India.
authors.forename: Narasimha
authors.initials: N
authors.lastname: Anaganti
authors.forename: Bhakti
authors.initials: B
authors.lastname: Basu
authors.forename: Shree Kumar
authors.initials: SK
authors.lastname: Apte
mesh_terms: D004249:DNA Damage / Q000528:radiation effects; D034301:Deinococcus / Q000235:genetics* / Q000528:radiation effects; D004926:Escherichia coli / Q000235:genetics; D005720:Gamma Rays; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics* / Q000528:radiation effects; D006801:Humans; D011401:Promoter Regions, Genetic / Q000528:radiation effects; D011836:Radiation Tolerance / Q000235:genetics*; D014158:Transcription, Genetic