Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0
<p>Abstract</p> <p>Background</p> <p>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...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2005-09-01
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| Series: | BMC Microbiology |
| Online Access: | http://www.biomedcentral.com/1471-2180/5/50 |
| _version_ | 1811314969721962496 |
|---|---|
| author | Wachocki Susi Prichard Kimberly Clary Susan Weyand Nathan J Howie Heather Jens Jason Keller Thomas J Langford Chris Potter Laura Friedrich Alexandra Mason Roxie M Du Ying Altherr Michael R Brettin Thomas Sodergren Erica Dillard Joseph P Weinstock George So Magdalene Arvidson Cindy |
| author_facet | Wachocki Susi Prichard Kimberly Clary Susan Weyand Nathan J Howie Heather Jens Jason Keller Thomas J Langford Chris Potter Laura Friedrich Alexandra Mason Roxie M Du Ying Altherr Michael R Brettin Thomas Sodergren Erica Dillard Joseph P Weinstock George So Magdalene Arvidson Cindy |
| author_sort | Wachocki Susi |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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, <it>Neisseria gonorrhoeae</it>, 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.</p> <p>Results</p> <p>The clone set thus far includes 1672 of the 2250 predicted ORFs of the <it>N. gonorrhoeae </it>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.</p> <p>Conclusion</p> <p>This archived set of Gateway<sup>® </sup>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.</p> |
| first_indexed | 2024-04-13T11:21:59Z |
| format | Article |
| id | doaj.art-3964f755957345edbf8118463c40c9b7 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-04-13T11:21:59Z |
| publishDate | 2005-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-3964f755957345edbf8118463c40c9b72022-12-22T02:48:47ZengBMCBMC Microbiology1471-21802005-09-01515010.1186/1471-2180-5-50Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0Wachocki SusiPrichard KimberlyClary SusanWeyand Nathan JHowie HeatherJens JasonKeller Thomas JLangford ChrisPotter LauraFriedrich AlexandraMason Roxie MDu YingAltherr Michael RBrettin ThomasSodergren EricaDillard Joseph PWeinstock GeorgeSo MagdaleneArvidson Cindy<p>Abstract</p> <p>Background</p> <p>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, <it>Neisseria gonorrhoeae</it>, 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.</p> <p>Results</p> <p>The clone set thus far includes 1672 of the 2250 predicted ORFs of the <it>N. gonorrhoeae </it>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.</p> <p>Conclusion</p> <p>This archived set of Gateway<sup>® </sup>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.</p>http://www.biomedcentral.com/1471-2180/5/50 |
| spellingShingle | Wachocki Susi Prichard Kimberly Clary Susan Weyand Nathan J Howie Heather Jens Jason Keller Thomas J Langford Chris Potter Laura Friedrich Alexandra Mason Roxie M Du Ying Altherr Michael R Brettin Thomas Sodergren Erica Dillard Joseph P Weinstock George So Magdalene Arvidson Cindy Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 BMC Microbiology |
| title | Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 |
| title_full | Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 |
| title_fullStr | Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 |
| title_full_unstemmed | Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 |
| title_short | Expression capable library for studies of <it>Neisseria gonorrhoeae</it>, version 1.0 |
| title_sort | expression capable library for studies of it neisseria gonorrhoeae it version 1 0 |
| url | http://www.biomedcentral.com/1471-2180/5/50 |
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