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Application of chromosomal DNA and protein targeting for the identification of Y ersinia pestis
Author(s) -
Larson Marilynn A.,
Ding ShiJian,
Slater Shawn R.,
Hanway Anna,
Bartling Amanda M.,
Fey Paul D.,
Lockridge Oksana,
Francesconi Stephen C.,
Hinrichs Steven H.
Publication year - 2013
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201200092
Subject(s) - yersinia pestis , biology , locus (genetics) , computational biology , proteomics , biomarker , polymerase chain reaction , genetics , virulence , microbiology and biotechnology , gene
Purpose A comprehensive strategy was developed and validated for the identification of pathogens from closely related near neighbors using both chromosomal and protein biomarkers, with emphasis on distinguishing Y ersinia pestis from the ancestral bacterium Y ersinia pseudotuberculosis . Experimental design Computational analysis was used to discover chromosomal targets unique to Y . pestis . Locus identifier YP O1670 was selected for further validation and PCR was used to confirm that this biomarker was exclusively present in Y . pestis strains, while absent in other Y ersinia species. RT ‐ PCR and W estern blot analyses were utilized to evaluate YP O1670 expression and MRM MS was performed to identify the YP O1670 protein within cell lysates. Results The described study validated that YP O1670 was exclusive to Y . pestis . PCR confirmed the locus to be unique to Y . pestis . The associated transcript and protein were produced throughout growth with the highest abundance occurring in stationary phase and MRM MS conclusively identified the YP O1670 protein in cell extracts. Conclusions and clinical relevance These findings validated YP O1670 as a reliable candidate biomarker for Y . pestis and that a dual DNA and protein targeting approach is feasible for the development of next‐generation assays to accurately differentiate pathogens from near neighbors.