In situ real-time evaluation of radiation-responsive promoters in the extremely radioresistant microbe Deinococcus radiodurans

Journal of biosciences

Shree Kumar Apte1¹  Bhakti Basu¹  Narasimha Anaganti¹ 

[1] Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India$$

关键词: Acclimation;    insect hormone;    longevity;    seasonal polyphenism;    starvation resistance;   

DOI  :  

来源: Indian Academy of Sciences

【 摘 要 】

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.

【 授权许可】

Unknown