dc.contributor.author | Nicastro, Jessica | |
dc.contributor.author | Sheldon, Katlyn | |
dc.contributor.author | El-zarkout, Farah A. | |
dc.contributor.author | Sokolenko, Stanislav | |
dc.contributor.author | Aucoin, Marc | |
dc.contributor.author | Slavcev, Roderick | |
dc.date.accessioned | 2017-06-05 14:28:43 (GMT) | |
dc.date.available | 2017-06-05 14:28:43 (GMT) | |
dc.date.issued | 2013-09-01 | |
dc.identifier.uri | http://dx.doi.org/10.1007/s00253-013-4898-6 | |
dc.identifier.uri | http://hdl.handle.net/10012/11986 | |
dc.description.abstract | The Bacteriophage lambda capsid protein gpD has been used extensively for fusion polypeptides that can be expressed from plasmids in Escherichia coli and remain soluble. In this study, a genetically controlled dual expression system for the display of enhanced green fluorescent protein (eGFP) was developed and characterized. Wild-type D protein (gpD) expression is encoded by lambda Dam15 infecting phage particles, which can only produce a functional gpD protein when translated in amber suppressor strains of E. coli in the absence of complementing gpD from a plasmid. However, the isogenic suppressors vary dramatically in their ability to restore functional packaging to lambda Dam15, imparting the first dimension of decorative control. In combination, the D-fusion protein, gpD::eGFP, was supplied in trans from a multicopy temperature-inducible expression plasmid, influencing D::eGFP expression and hence the availability of gpD::eGFP to complement for the Dam15 mutation and decorate viable phage progeny. Despite being the worst suppressor, maximal incorporation of gpD::eGFP into the lambda Dam15 phage capsid was imparted by the SupD strain, conferring a gpDQ68S substitution, induced for plasmid expression of pD::eGFP. Differences in size, fluorescence and absolute protein decoration between phage preparations could be achieved by varying the temperature of and the suppressor host carrying the pD::eGFP plasmid. The effective preparation with these two variables provides a simple means by which to manage fusion decoration on the surface of phage lambda. | en |
dc.description.sponsorship | UW Start-up funds; Drug Safety and Effectiveness Cross-Disciplinary Training (DSECT) Scholarship; Canadian Institute of Health Research (CIHR) | en |
dc.language.iso | en | en |
dc.publisher | Springer | en |
dc.rights | Attribution 3.0 Unported | * |
dc.rights.uri | https://creativecommons.org/licenses/by/3.0/ | * |
dc.subject | Recombinant Escherichia-Coli | en |
dc.subject | Cell-Density Cultures | en |
dc.subject | Bacteriophage-Lambda | en |
dc.subject | Transcriptional Response | en |
dc.subject | Insecticidal Toxin | en |
dc.subject | Expression System | en |
dc.subject | Efficient Display | en |
dc.subject | CDNA Libraries | en |
dc.subject | Proteins | en |
dc.subject | Surface | en |
dc.title | Construction and analysis of a genetically tuneable lytic phage display system | en |
dc.type | Article | en |
dcterms.bibliographicCitation | Nicastro, J., Sheldon, K., El-zarkout, F. A., Sokolenko, S., Aucoin, M. G., & Slavcev, R. (2013). Construction and analysis of a genetically tuneable lytic phage display system. Applied Microbiology and Biotechnology, 97(17), 7791–7804. https://doi.org/10.1007/s00253-013-4898-6 | en |
uws.contributor.affiliation1 | Faculty of Engineering | en |
uws.contributor.affiliation2 | Chemical Engineering | en |
uws.typeOfResource | Text | en |
uws.typeOfResource | Text | en |
uws.peerReviewStatus | Reviewed | en |
uws.scholarLevel | Faculty | en |