NDLI: Construction of recombinant Nitrosomonas europaea expressing green fluorescent protein in response to co-oxidation of chloroform

Content Provider	Springer Nature Link
Author	Gvakharia, Barbara O. Bottomley, Peter J. Arp, Daniel J. Sayavedra Soto, Luis A.
Copyright Year	2009
Abstract	Transcriptional fusions with gfp driven by the promoter region of mbla (NE2571) in pPRO/mbla4 and clpB (NE2402) in pPRO/clpb7 were used to transform the ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718). The two genes were chosen because their transcript levels were found at much higher levels in N. europaea in response to oxidation of chloroform and chloromethane. In N. europaea transformed with pPRO/mbla4, green fluorescent protein (GFP)-dependent fluorescence increased from 3- to 18-fold above control levels in response to increasing chloroform concentrations (7 to 28 μM), and from 8- to 10-fold in response to increasing hydrogen peroxide concentrations (2.5–7.5 mM). The GFP-dependent fluorescence of N. europaea transformed with pPRO/clpb7 also showed an increase of 6- to 10-fold in response to chloroform (28–100 μM) but did not respond to H2O2. Our data provide proof of concept that biosensors can be fabricated in ammonia-oxidizing bacteria using “sentinel” genes that up-regulate in response to stress caused either by co-oxidation of chlorinated solvents or by the presence of H2O2. The fabricated biosensors had a consistent concentration-dependent response to chloroform; however, these did not respond to other chlorinated compounds that cause similar cellular stress.
Starting Page	1179
Ending Page	1185
Page Count	7
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	82
Issue Number	6
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2009-02-27
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Nitrosomonas europaea Biosensor Green fluorescent protein Chloroform Hydrogen peroxide Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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