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Expression capable library for studies of Neisseria gonorrhoeae, version 1.0

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Brettin, Thomas S. (Thomas Scott), Altherr, Michael R., Du, Ying, Mason, Roxie M., Friedrich, Alexandra, Potter, Laura, Langford, Chris, Keller, Thomas J., Jens, Jason, Howie, Heather, Weyand, Nathan J., Clary, Susan, Prichard, Kimberley, Wachocki, Susi, Sodergren, Erica, Dillard, Joseph P., Dillard, George, So, Magdalene and Grove Arvidson , Cindy. (2005) Expression capable library for studies of Neisseria gonorrhoeae, version 1.0. BMC Microbiology, Vol.5 (No.50). ISSN 1471-2180

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Official URL: http://dx.doi.org/10.1186/1471-2180-5-50

Abstract

Background The sexually transmitted disease, gonorrhea, is a serious health problem in developed as well as in developing countries, for which treatment continues to be a challenge. The recent completion of the genome sequence of the causative agent, Neisseria gonorrhoeae, opens up an entirely new set of approaches for studying this organism and the diseases it causes. Here, we describe the initial phases of the construction of an expression-capable clone set representing the protein-coding ORFs of the gonococcal genome using a recombination-based cloning system. Results The clone set thus far includes 1672 of the 2250 predicted ORFs of the N. gonorrhoeae genome, of which 1393 (83%) are sequence-validated. Included in this set are 48 of the 61 ORFs of the gonococcal genetic island of strain MS11, not present in the sequenced genome of strain FA1090. L-arabinose-inducible glutathione-S-transferase (GST)-fusions were constructed from random clones and each was shown to express a fusion protein of the predicted size following induction, demonstrating the use of the recombination cloning system. PCR amplicons of each ORF used in the cloning reactions were spotted onto glass slides to produce DNA microarrays representing 2035 genes of the gonococcal genome. Pilot experiments indicate that these arrays are suitable for the analysis of global gene expression in gonococci. Conclusion This archived set of Gateway® entry clones will facilitate high-throughput genomic and proteomic studies of gonococcal genes using a variety of expression and analysis systems. In addition, the DNA arrays produced will allow us to generate gene expression profiles of gonococci grown in a wide variety of conditions. Together, the resources produced in this work will facilitate experiments to dissect the molecular mechanisms of gonococcal pathogenesis on a global scale, and ultimately lead to the determination of the functions of unknown genes in the genome.

Item Type:	Journal Article
Subjects:	R Medicine > R Medicine (General) Q Science > QR Microbiology
Divisions:	Faculty of Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	Neisseria gonorrhea, Gene mapping
Journal or Publication Title:	BMC Microbiology
Publisher:	BioMed Central Ltd.
ISSN:	1471-2180
Date:	1 September 2005
Volume:	Vol.5
Number:	No.50
Identification Number:	10.1186/1471-2180-5-50
Status:	Peer Reviewed
Access rights to Published version:	Open Access
Funder:	Michigan State University. College of Human Medicine, Michigan State University. College of Osteopathic Medicine, Michigan State University. Vice President for Research and Graduate Studies, United States. Dept. of Energy, Los Alamos National Laboratory (LANL), National Institutes of Health (U.S.) (NIH), Deutscher Akademischer Austauschdienst (DAAD)
Grant number:	Intramural research grant (MSU VP), Molecular Foundations of Pathogenesis project (W-7405- ENG-36) (DofE), AI34560, AI47260 and AI49973 (NIH),
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URI:	http://wrap.warwick.ac.uk/id/eprint/581