Expression capable library for studies of Neisseria gonorrhoeae, version 1.0

Thomas Brettin, Michael R. Altherr, Ying Du, Roxie M. Mason, Alexandra Friedrich, Laura Potter, Chris Langford, Thomas J. Keller, Jason Jens, Heather Howie, Nathan J. Weyand, Susan Clary, Kimberly Prichard, Susi Wachocki, Erica Sodergren, Joseph P. Dillard, George Weinstock, Magdalene "Maggie" So, Cindy Grove Arvidson

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish (US)
Article number50
JournalBMC Microbiology
Volume5
DOIs
StatePublished - Sep 1 2005
Externally publishedYes

Fingerprint

Neisseria gonorrhoeae
Libraries
Genome
Open Reading Frames
Clone Cells
Organism Cloning
Oligonucleotide Array Sequence Analysis
Genetic Recombination
Genes
Arabinose
Gonorrhea
Sexually Transmitted Diseases
Glutathione Transferase
Transcriptome
Proteomics
Developing Countries
Glass
Proteins
Gene Expression
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Brettin, T., Altherr, M. R., Du, Y., Mason, R. M., Friedrich, A., Potter, L., ... Arvidson, C. G. (2005). Expression capable library for studies of Neisseria gonorrhoeae, version 1.0. BMC Microbiology, 5, [50]. https://doi.org/10.1186/1471-2180-5-50

Expression capable library for studies of Neisseria gonorrhoeae, version 1.0. / Brettin, Thomas; 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, Kimberly; Wachocki, Susi; Sodergren, Erica; Dillard, Joseph P.; Weinstock, George; So, Magdalene "Maggie"; Arvidson, Cindy Grove.

In: BMC Microbiology, Vol. 5, 50, 01.09.2005.

Research output: Contribution to journalArticle

Brettin, T, Altherr, MR, Du, Y, Mason, RM, Friedrich, A, Potter, L, Langford, C, Keller, TJ, Jens, J, Howie, H, Weyand, NJ, Clary, S, Prichard, K, Wachocki, S, Sodergren, E, Dillard, JP, Weinstock, G, So, MM & Arvidson, CG 2005, 'Expression capable library for studies of Neisseria gonorrhoeae, version 1.0', BMC Microbiology, vol. 5, 50. https://doi.org/10.1186/1471-2180-5-50
Brettin T, Altherr MR, Du Y, Mason RM, Friedrich A, Potter L et al. Expression capable library for studies of Neisseria gonorrhoeae, version 1.0. BMC Microbiology. 2005 Sep 1;5. 50. https://doi.org/10.1186/1471-2180-5-50
Brettin, Thomas ; 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, Kimberly ; Wachocki, Susi ; Sodergren, Erica ; Dillard, Joseph P. ; Weinstock, George ; So, Magdalene "Maggie" ; Arvidson, Cindy Grove. / Expression capable library for studies of Neisseria gonorrhoeae, version 1.0. In: BMC Microbiology. 2005 ; Vol. 5.
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