Abstract：
:When studying interspecies interactions in a bacterial consortium, it may be desirable to analyze one species' transcriptional response as influenced by the other species. We developed a dual fluorescence system of Escherichia coli for Fluorescence-Activated Cell Sorter (FACS)-based analysis for such a purpose. First, we generated E. coli SCC1 strain, which constitutively expresses green fluorescent protein (GFP), but otherwise showed no observable difference from the parent strain MG1655 with respect to morphology, growth, and FACS-analyzed side- and forward-scatter profiles. Next, to analyze transcriptional response, plasmids carrying promoters of interest fused to a red fluorescent protein (AsRed2) reporter, were introduced into strain SCC1. Quantification of promoter activities of araB, lacZ, fadB and rpoE via AsRed2 reporter verified that the induction levels are similar between MG1655 and SCC1 strains. In mixtures and co-cultures, GFP expression of E. coli SCC1 allowed it to be separated from non-E. coli species by FACS to purity levels of 96.7-100.0%. When a mixture of E. coli SCC1 carrying promoter-AsRed2 fusion and a non-E. coli strain was analyzed by FACS, it enabled (i) distinction of E. coli SCC1 from the non-E. coli strain, (ii) analysis of the E. coli promoter activity via AsRed2 expression and (iii) identification of transcriptional heterogeneity within the E. coli population. Co-cultures of E. coli SCC1 with Klebsiella pneumoniae and/or Enterococcus faecalis analyzed by FACS showed that E. coli fadB and rpoE transcription were differentially influenced by partner species.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Miao H,Ratnasingam S,Pu CS,Desai MM,Sze CCdoi
10.1016/j.mimet.2008.09.015subject
Has Abstractpub_date
2009-02-01 00:00:00pages
109-19issue
2eissn
0167-7012issn
1872-8359pii
S0167-7012(08)00345-Xjournal_volume
76pub_type
杂志文章abstract：:Despite advances in antifungal prophylaxis and therapy, morbidity and mortality incurred by yeasts remain a significant burden. As pathogenic yeast species vary in their susceptibilities to antifungal agents, clinical microbiology laboratories face an important challenge to identify them rapidly and accurately. Althou...
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journal_title：Journal of microbiological methods
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journal_title：Journal of microbiological methods
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journal_title：Journal of microbiological methods
pub_type： 杂志文章
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journal_title：Journal of microbiological methods
pub_type： 杂志文章,评审
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journal_title：Journal of microbiological methods
pub_type： 杂志文章
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journal_title：Journal of microbiological methods
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journal_title：Journal of microbiological methods
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