Determining the repertoire of immunodominant proteins via whole-genome amplification of intracellular pathogens.

PloS One
Michael J DarkAnthony F Barbet

Abstract

Culturing many obligate intracellular bacteria is difficult or impossible. However, these organisms have numerous adaptations allowing for infection persistence and immune system evasion, making them some of the most interesting to study. Recent advancements in genome sequencing, pyrosequencing and Phi29 amplification, have allowed for examination of whole-genome sequences of intracellular bacteria without culture. We have applied both techniques to the model obligate intracellular pathogen Anaplasma marginale and the human pathogen Anaplasma phagocytophilum, in order to examine the ability of phi29 amplification to determine the sequence of genes allowing for immune system evasion and long-term persistence in the host. When compared to traditional pyrosequencing, phi29-mediated genome amplification had similar genome coverage, with no additional gaps in coverage. Additionally, all msp2 functional pseudogenes from two strains of A. marginale were detected and extracted from the phi29-amplified genomes, highlighting its utility in determining the full complement of genes involved in immune evasion.

References

Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Jan 1, 1996·Emerging Infectious Diseases·D H Walker, J S Dumler
Oct 18, 2000·Infection and Immunity·A F BarbetG H Palmer
Mar 29, 2002·Molecular Microbiology·Kelly A BraytonAnthony F Barbet
Jan 22, 2003·Molecular Microbiology·Patrick F M MeeusAnthony F Barbet
Dec 25, 2004·Proceedings of the National Academy of Sciences of the United States of America·Kelly A BraytonDonald P Knowles
Dec 2, 2005·Current Opinion in Hematology·Jason A Carlyon, Erol Fikrig
Feb 17, 2006·PLoS Genetics·Julie C Dunning HotoppHervé Tettelin
Nov 2, 2006·BMC Biotechnology·Roderick F FelsheimUlrike G Munderloh
Jan 1, 2008·Nature Methods·Stephan C Schuster
Feb 7, 2008·Proceedings of the National Academy of Sciences of the United States of America·James E FutseGuy H Palmer
Sep 9, 2008·Genome Research·Douglas R SmithPaul M Richardson
Mar 25, 2011·Methods in Molecular Biology·Yuichi Hongoh, Atsushi Toyoda
Jul 8, 2011·The American Journal of Tropical Medicine and Hygiene·F Scott DahlgrenJennifer H McQuiston
Nov 10, 2011·FEMS Immunology and Medical Microbiology·Guy H PalmerKelly A Brayton

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Citations

Oct 16, 2013·Current Microbiology·Pablo A NuñezMarisa D Farber
Jan 7, 2016·Annual Review of Animal Biosciences·Wendy C Brown, Anthony F Barbet
Sep 1, 2015·Frontiers in Cellular and Infection Microbiology·Thibaud DugatNadia Haddad
Sep 10, 2016·BioMed Research International·Rosa Estela Quiroz-CastañedaSergio Darío Rodríguez-Camarillo
Jan 26, 2017·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Mara BattilaniFrancesco Dondi

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Datasets Mentioned

BETA
SRA050330.2
CP000235

Software Mentioned

Artemis
Mafft
Mosaik
CLC Genomics Workbench
Jalview
samtools
custom Perl program
Se
Al
Newbler

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