scholarly journals Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages

2020 ◽  
Vol 8 (6) ◽  
pp. 934 ◽  
Author(s):  
Peter Braun ◽  
Immanuel Wolfschläger ◽  
Leonie Reetz ◽  
Lilia Bachstein ◽  
Ana Clara Jacinto ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Anthracis ◽  
Receptor Binding ◽  
Fusion Proteins ◽  
Binding Proteins ◽  
Fluorescent Proteins ◽  
Lambdoid Prophage ◽  
Unambiguous Detection ◽  
Close Genetic Relationship ◽  
Bacillus Cereus Sensu Lato

Bacillus anthracis, the etiological agent of anthrax disease, is typically diagnosed by immunological and molecular methods such as polymerase chain reaction (PCR). Alternatively, mass spectrometry techniques may aid in confirming the presence of the pathogen or its toxins. However, because of the close genetic relationship between B. anthracis and other members of the Bacillus cereus sensu lato group (such as Bacillus cereus or Bacillus thuringiensis) mis- or questionable identification occurs frequently. Also, bacteriophages such as phage gamma (which is highly specific for B. anthracis) have been in use for anthrax diagnostics for many decades. Here we employed host cell-specific receptor binding proteins (RBP) of (pro)-phages, also known as tail or head fibers, to develop a microscopy-based approach for the facile, rapid and unambiguous detection of B. anthracis cells. For this, the genes of (putative) RBP from Bacillus phages gamma, Wip1, AP50c and from lambdoid prophage 03 located on the chromosome of B. anthracis were selected. Respective phage genes were heterologously expressed in Escherichia coli and purified as fusions with fluorescent proteins. B. anthracis cells incubated with either of the reporter fusion proteins were successfully surface-labeled. Binding specificity was confirmed as RBP fusion proteins did not bind to most isolates of a panel of other B. cereus s.l. species or to more distantly related bacteria. Remarkably, RBP fusions detected encapsulated B. anthracis cells, thus RBP were able to penetrate the poly-γ-d-glutamate capsule of B. anthracis. From these results we anticipate this RBP-reporter assay may be useful for rapid confirmative identification of B. anthracis.

scholarly journals Genomic characterization of the Bacillus cereus sensu lato species: Backdrop to the evolution of Bacillus anthracis

2012 ◽  
Vol 22 (8) ◽  
pp. 1512-1524 ◽  
Author(s):  
M. E. Zwick ◽  
S. J. Joseph ◽  
X. Didelot ◽  
P. E. Chen ◽  
K. A. Bishop-Lilly ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Anthracis ◽  
Genomic Characterization ◽  
Bacillus Cereus Sensu Lato

scholarly journals Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1462
Author(s):  
Peter Braun ◽  
Nadja Rupprich ◽  
Diana Neif ◽  
Gregor Grass
Keyword(s):  
Bacillus Anthracis ◽  
Receptor Binding ◽  
Pathogen Detection ◽  
Fluorescent Proteins ◽  
Binding Protein ◽  
Host Cells ◽  
Close Relationship ◽  
Phage Receptor ◽  
Protein Assays ◽  
Receptor Binding Protein

Bacteriophage receptor binding proteins (RBPs) are employed by viruses to recognize specific surface structures on bacterial host cells. Recombinant RBPs have been utilized for detection of several pathogens, typically as fusions with reporter enzymes or fluorescent proteins. Identification of Bacillus anthracis, the etiological agent of anthrax, can be difficult because of the bacterium’s close relationship with other species of the Bacillus cereussensu lato group. Here, we facilitated the identification of B. anthracis using two implementations of enzyme-linked phage receptor binding protein assays (ELPRA). We developed a single-tube centrifugation assay simplifying the rapid analysis of suspect colonies. A second assay enables identification of suspect colonies from mixed overgrown solid (agar) media derived from the complex matrix soil. Thus, these tests identified vegetative cells of B. anthracis with little processing time and may support or confirm pathogen detection by molecular methods such as polymerase chain reaction.

scholarly journals Draft Genome Sequences from a Novel Clade of Bacillus cereus Sensu Lato Strains, Isolated from the International Space Station

2017 ◽  
Vol 5 (32) ◽  
Author(s):  
Kasthuri Venkateswaran ◽  
Aleksandra Checinska Sielaff ◽  
Shashikala Ratnayake ◽  
Robert K. Pope ◽  
Thomas E. Blank ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Bacillus Anthracis ◽  
International Space Station ◽  
Draft Genome ◽  
Space Station ◽  
Genome Sequences ◽  
Content Type ◽  
International Space ◽  
Experiment Module ◽  
Bacillus Cereus Sensu Lato

ABSTRACT The draft genome sequences of six Bacillus strains, isolated from the International Space Station and belonging to the Bacillus anthracis-B. cereus-B. thuringiensis group, are presented here. These strains were isolated from the Japanese Experiment Module (one strain), U.S. Harmony Node 2 (three strains), and Russian Segment Zvezda Module (two strains).

scholarly journals A Recombinant Carboxy-Terminal Domain of the Protective Antigen of Bacillus anthracis Protects Mice against Anthrax Infection

2002 ◽  
Vol 70 (3) ◽  
pp. 1653-1656 ◽  
Author(s):  
Helen C. Flick-Smith ◽  
Nicola J. Walker ◽  
Paula Gibson ◽  
Helen Bullifent ◽  
Sarah Hayward ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Fusion Proteins ◽  
Protective Antigen ◽  
Protective Efficacy ◽  
Terminal Domain ◽  
Carboxy Terminal Domain ◽  
Anthrax Infection ◽  
Carboxy Terminal ◽  
Domain 4

ABSTRACT The immunogenicity and protective efficacy of overlapping regions of the protective antigen (PA) polypeptide, cloned and expressed as glutathione S-transferase fusion proteins, have been assessed. Results show that protection can be attributed to individual domains and imply that it is domain 4 which contains the dominant protective epitopes of PA.

scholarly journals Impact of fluorescent protein fusions on the bacterial flagellar motor

2017 ◽  
Author(s):  
M Heo ◽  
AL Nord ◽  
D Chamousset ◽  
E van Rijn ◽  
HJE Beaumont ◽  
...  
Keyword(s):  
Side Effects ◽  
Protein Function ◽  
Fusion Proteins ◽  
Fluorescent Proteins ◽  
Protein Complexes ◽  
Switching Frequency ◽  
Flagellar Motor ◽  
Internal Dynamics ◽  
Biologically Relevant ◽  
Bacterial Flagellar Motor

AbstractFluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side effects. Such insight, however, is still lacking for many applications of fluorescent fusions. This is particularly relevant in the study of the internal dynamics of motor protein complexes, where both the chemical and mechanical reaction coordinates can be affected. Fluorescent proteins fused to thestatorof the bacterial flagellar motor (BFM) complex have previously been used to successfully unveil the internal subunit dynamics of the motor. Here we report the effects of three different fluorescent proteins fused to the stator, all of which altered BFM behavior. The torque generated by individual stators was reduced while their stoichiometry in the complex remained unaffected. MotB fusions decreased the rotation-direction switching frequency of single motors and induced a novel BFM behavior: a bias-dependent asymmetry in the speed attained in the two rotation directions. All these effects could be mitigated by the insertion of a linker at the fusion point. These findings provide a quantitative account of the effects of fluorescent fusions on BFM dynamics and their alleviation—new insights that advance the use of fluorescent fusions to probe the dynamics of protein complexes.Author summaryMuch of what is known about the biology of proteins was discovered by fusing them to fluorescent proteins that allow detection of their location. But the label comes at a cost: the presence of the tag can alter the behavior of the protein of interest in unforeseen, yet biologically relevant ways. These side effects limit the depth to which fluorescent proteins can be used to probe protein function. One of the systems that has been successfully studied with fluorescent fusions for which these effects have not been addressed are dynamic protein complexes that carry out mechanical work. We examined how fluorescent proteins fused to a component of the bacterial flagellar motor complex impacts its function. Our findings show that the fusion proteins altered biologically relevant dynamical properties of the motor, including induction of a novel mechanical behavior, and demonstrate an approach to alleviate this. These results advance our ability to dissect the bacterial flagellar motor, and the internal dynamics of protein complexes in general, with fluorescent fusion proteins while causing minimal perturbation.

scholarly journals Detection of Penicillin-Binding Proteins in Bacillus Cereus by Using Biotinylated β-Lactams

10.5109/24332 ◽  
2000 ◽  
Vol 44 (3/4) ◽  
pp. 299-307
Author(s):  
Takahisa Miyamoto ◽  
Kouji Sukimoto ◽  
Md Abu Sayed ◽  
Sam-In Kim ◽  
Ken-ichi Honjoh ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Binding Proteins ◽  
Penicillin Binding Proteins
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