Abstract
Methods aimed at identification of immune evasion proteins are mainly rely on in silico prediction of sequence, structural homology to known evasion proteins or use a proteomics driven approach. Although proven successful these methods are limited by a low efficiency and or lack of functional identification. Here we describe a high-throughput genomic strategy to functionally identify bacterial immune evasion proteins using phage display technology. Genomic bacterial DNA is randomly fragmented and ligated into a phage display vector that is used to create a phage display library expressing bacterial secreted and membrane bound proteins. This library is used to select displayed bacterial secretome proteins that interact with host immune components.
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References
Jongerius I, Köhl J, Pandey MK, Ruyken M, van Kessel KPM, van Strijp JAG et al (2007) Staphylococcal complement evasion by various convertase-blocking molecules. J Exp Med 204:2461–2471. doi:10.1084/jem.20070818
de Haas CJ, Veldkamp KE, Peschel A, Weerkamp F, Van Wamel WJ, Heezius EC et al (2004) Chemotaxis inhibitory protein of Staphylococcus aureus, a bacterial antiinflammatory agent. J Exp Med 199:687–695
Jankovic D, Collett MA, Lubbers MW, Rakonjac J (2007) Direct selection and phage display of a Gram-positive secretome. Genome Biol 8:R266. doi:10.1186/gb-2007-8-12-r266
Carlos F, Barbas III, Burton DR, Scott JK, Silverman GJ (2001) Phage display: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Hoogenboom HR (2002) Overview of antibody phage-display technology and its applications. Methods Mol Biol 178:1–37, doi:11968478
Karlström Å, Jacobsson K, Flock M, Flock J-I, Guss B (2004) Identification of a novel collagen-like protein, SclC, in Streptococcus equi using signal sequence phage display. Vet Microbiol 104:179–188. doi:10.1016/j.vetmic.2004.09.014
Fevre C, Bestebroer J, Mebius MM, de Haas CJC, van Strijp JAG, Fitzgerald JR et al (2014) Staphylococcus aureus proteins SSL6 and SElX interact with neutrophil receptors as identified using secretome phage display. Cell Microbiol 16:1646–1665. doi:10.1111/cmi.12313
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Fevre, C., Scheepmaker, L., Haas, PJ. (2017). Identifying Bacterial Immune Evasion Proteins Using Phage Display. In: Nordenfelt, P., Collin, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology, vol 1535. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6673-8_4
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DOI: https://doi.org/10.1007/978-1-4939-6673-8_4
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6673-8
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