Proximity ligation assays with peptide conjugate 'burrs' for the sensitive detection of spores.
The proximity ligation assay (PLA) has previously been used for the sensitive and specific detection of single proteins. In order to adapt PLA methods for the detection of cell surfaces, we have generated multivalent peptide-oligonucleotide-phycoerythrin conjugates ('burrs') that can bind adjacent to one another on a cell surface and be ligated together to form unique amplicons. Real-time PCR detection of burr ligation events specifically identified as few as 100 Bacillus anthracis, 10 Bacillus subtilis and 1 Bacillus cereus spore. Burrs should prove to be generally useful for detecting and mapping interactions and distances between cell surface proteins.
Pai
Supriya
S
Section of Molecular Genetics and Microbiology, School of Biological Sciences, University of Texas at Austin, Austin, TX 78712, USA.
Ellington
Andrew D
AD
Levy
Matthew
M
D001408:Bacillus anthracis / Q000302:isolation & purification / Q000502:physiology; D001409:Bacillus cereus / Q000302:isolation & purification / Q000502:physiology; D001412:Bacillus subtilis / Q000302:isolation & purification / Q000502:physiology; D011088:DNA Ligases; D015342:DNA Probes / Q000737:chemistry; D008856:Microscopy, Fluorescence; D015345:Oligonucleotide Probes / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010799:Phycoerythrin / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D013171:Spores, Bacterial / Q000302:isolation & purification*
D015342:DNA Probes; D015345:Oligonucleotide Probes; D010455:Peptides; D010799:Phycoerythrin; D011088:DNA Ligases