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An examination of an Antarctic soil metagenomic-derivate putative methylthioadenosine phosphorylase gene as a novel reporter gene for promoter trapping
Abstract
The rsfp gene was discovered during examination ofan Antarctic soil-derived metagenome library and encodesa bacterial methylthioadenosine phosphorylase (GenBank accession number GQ202582). In a previous study, we found that E. coli colonies expressing the rsfp gene reveal strong pink fluorescence after exposure to UV light when grown on culture medium supplemented with rhodamine B (Cieśliński et al., 2009). In contrast to that, the colonies of the E. coli strains, defi cient in the rsfp gene, do not show any fluorescence under the same experimental conditions.We found that the reason for the fl uorescence phenomenon of E. coli cells expressing the rsfp gene is the binding of the fluorescent dye rhodamine B (abbrev. RB) molecules on RSFP protein molecules thatleads to the increase in the fluorescence yield of RB molecules excited by UV light and the accumulation of RB molecules in E. coli cells during colony growth (manuscript in preparation). Based on these results we decided to examine the potential of the rsfp gene as a novel reporter gene for identifying the location of the putative promoter sequences in the E. coli TOP10F' genome fragments cloned to a pTrap vector constructed in this work. The E. coli TOP10F' strain was used as model organism in this study. On the other hand, because of the expression of the rsfp gene in the E. coli cells leads to the accumulation of the rhodamine B that is the fluorescent dye, we also examined our new RSFP-RB reporter system by fluorescence microscopy.
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
-
JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY
no. 58,
pages 387 - 395,
ISSN: 0022-1260 - Language:
- English
- Publication year:
- 2012
- Bibliographic description:
- Bartasun P., Cieśliński H., Kur J.: An examination of an Antarctic soil metagenomic-derivate putative methylthioadenosine phosphorylase gene as a novel reporter gene for promoter trapping// JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY. -Vol. 58, nr. iss. 5 (2012), s.387-395
- DOI:
- Digital Object Identifier (open in new tab) 10.2323/jgam.58.387
- Verified by:
- Gdańsk University of Technology
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