scholarly journals Improvement of the Fluorescence Intensity during a Flow Cytometric Analysis for Rice Protoplasts by Localization of a Green Fluorescent Protein into Chloroplasts

2014 ◽  
Vol 16 (1) ◽  
pp. 788-804 ◽  
Author(s):  
Min You ◽  
Sun-Hyung Lim ◽  
Min-Jin Kim ◽  
Ye Jeong ◽  
Mi-Gi Lee ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescence Intensity ◽  
Fluorescent Protein ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Green Fluorescent

scholarly journals Transformation of Rickettsia prowazekii to Erythromycin Resistance Encoded by the Escherichia coli ereB Gene

2000 ◽  
Vol 182 (11) ◽  
pp. 3289-3291 ◽  
Author(s):  
Lyudmila I. Rachek ◽  
Andria Hines ◽  
Aimee M. Tucker ◽  
Herbert H. Winkler ◽  
David O. Wood
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Flow Cytometric Analysis ◽  
Etiologic Agent ◽  
Erythromycin Resistance ◽  
Rickettsia Prowazekii ◽  
Flow Cytometric ◽  
Epidemic Typhus ◽  
Green Fluorescent

ABSTRACT Rickettsia prowazekii, the etiologic agent of epidemic typhus, is an obligate, intracytoplasmic, parasitic bacterium. Recently, the transformation of this bacterium via electroporation has been reported. However, in these studies identification of transformants was dependent upon either selection of an R. prowazekii rpoB chromosomal mutation imparting rifampin resistance or expression of the green fluorescent protein and flow cytometric analysis. In this paper we describe the expression inR. prowazekii of the Escherichia coli ereBgene. This gene codes for an erythromycin esterase that cleaves erythromycin. To the best of our knowledge, this is the first report of the expression of a nonrickettsial, antibiotic-selectable gene inR. prowazekii. The availability of a positive selection for rickettsial transformants is an important step in the characterization of genetic analysis systems in the rickettsiae.

A Flow Cytometric Protocol for Titering Recombinant Adenoviral Vectors Containing the Green Fluorescent Protein

2000 ◽  
Vol 14 (3) ◽  
pp. 197-203 ◽  
Author(s):  
Daron C. Hitt ◽  
J. Leland Booth ◽  
Viji Dandapani ◽  
Larry R. Pennington ◽  
Jeffrey M. Gimble ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Adenoviral Vectors ◽  
Flow Cytometric ◽  
Green Fluorescent

scholarly journals Use of Green Fluorescent Protein To Assess Urease Gene Expression by Uropathogenic Proteus mirabilis during Experimental Ascending Urinary Tract Infection

1998 ◽  
Vol 66 (1) ◽  
pp. 330-335 ◽  
Author(s):  
Hui Zhao ◽  
Richard B. Thompson ◽  
Virginia Lockatell ◽  
David E. Johnson ◽  
Harry L. T. Mobley
Keyword(s):  
Gene Expression ◽  
Urinary Tract Infection ◽  
Green Fluorescent Protein ◽  
Urinary Tract ◽  
Tract Infection ◽  
Fluorescence Intensity ◽  
Proteus Mirabilis ◽  
Fluorescent Protein ◽  
Green Fluorescent

ABSTRACT Proteus mirabilis, a cause of complicated urinary tract infection, expresses urease when exposed to urea. While it is recognized that the positive transcriptional activator UreR induces gene expression, the levels of expression of the enzyme during experimental infection are not known. To investigate in vivo expression of P. mirabilis urease, the gene encoding green fluorescent protein (GFP) was used to construct reporter fusions. Translational fusions of urease accessory gene ureD, which is preceded by a urea-inducible promoter, were made withgfp (modified to express S65T/V68L/S72A [B. P. Cormack et al. Gene 173:33–38, 1996]). Constructs were confirmed by sequencing of the fusion junctions. UreD-GFP fusion protein was induced by urea in both Escherichia coli DH5α and P. mirabilis HI4320. By using Western blotting with antiserum raised against GFP, expression level was shown to correlate with urea concentration (tested from 0 to 500 mM), with highest induction at 200 to 500 mM urea. Fluorescent E. coli and P. mirabilis bacteria were observed by fluorescence microscopy following urea induction, and the fluorescence intensity of GFP in cell lysates was measured by spectrophotofluorimetry. P. mirabilis HI4320 carrying the UreD-GFP fusion plasmid was transurethrally inoculated into the bladders of CBA mice. One week postchallenge, fluorescent bacteria were detected in thin sections of both bladder and kidney samples; the fluorescence intensity of bacteria in bladder tissue was higher than that in the kidney. Kidneys were primarily infected with single-cell-form fluorescent bacteria, while aggregated bacterial clusters were observed in the bladder. Elongated swarmer cells were only rarely observed. These observations demonstrate that urease is expressed in vivo and that using GFP as a reporter protein is a viable approach to investigate in vivo expression ofP. mirabilis virulence genes in experimental urinary tract infection.

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