Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (833K) Freely available
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (60)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Thanassi, J. A.
Right arrow Articles by Pucci, M. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Thanassi, J. A.
Right arrow Articles by Pucci, M. J.
Related Collections
Right arrow Genomics
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2002, Vol. 30, No. 14 3152-3162
© 2002 Oxford University Press

Identification of 113 conserved essential genes using a high-throughput gene disruption system in Streptococcus pneumoniae

Jane A. Thanassi, Sandra L. Hartman-Neumann, Thomas J. Dougherty, Brian A. Dougherty1 and Michael J. Pucci*

Department of Microbiology and 1 Department of Applied Genomics, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, CT 06492, USA

*To whom correspondence should be addressed at present address: Achillion Pharmaceuticals, 300 George Street, New Haven, CT 06511, USA. Tel: +1 203 624 7000; Fax: +1 203 624 7003; Email: mpucci{at}achillion.com
Present addresses:
Jane A. Thanassi, Achillion Pharmaceuticals, 300 George Street, New Haven, CT 06511, USA
Sandra L. Hartman-Neumann and Thomas J. Dougherty, AI2, Pfizer Global Research and Development, Mailstop 8220-2358, Eastern Point Road, Groton, CT 06340, USA

The recent availability of bacterial genome sequence information permits the identification of conserved genes that are potential targets for novel antibiotic drug discovery. Using a coupled bioinformatic/experimental approach, a list of candidate conserved genes was generated using a Microbial Concordance bioinformatics tool followed by a targeted disruption campaign. Pneumococcal sequence data allowed for the design of precise PCR primers to clone the desired gene target fragments into the pEVP3 ‘suicide vector’. An insertion–duplication approach was employed that used the pEVP3 constructs and resulted in the introduction of a selectable chloramphenicol resistance marker into the chromosome. In the case of non-essential genes, cells can survive the disruption and form chloramphenicol-resistant colonies. A total of 347 candidate reading frames were subjected to disruption analysis, with 113 presumed to be essential due to lack of recovery of antibiotic-resistant colonies. In addition to essentiality determination, the same high-throughput methodology was used to overexpress gene products and to examine possible polarity effects for all essential genes.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Appl. Environ. Microbiol.Home page
J. C. Zweers, T. Wiegert, and J. M. van Dijl
Stress-Responsive Systems Set Specific Limits to the Overproduction of Membrane Proteins in Bacillus subtilis
Appl. Envir. Microbiol., December 1, 2009; 75(23): 7356 - 7364.
[Abstract] [Full Text] [PDF]

Home page
 Antimicrob. Agents Chemother.Home page
N. Ruiz
Streptococcus pyogenes YtgP (Spy_0390) Complements Escherichia coli Strains Depleted of the Putative Peptidoglycan Flippase MurJ
Antimicrob. Agents Chemother., August 1, 2009; 53(8): 3604 - 3605.
[Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. I. Handford, B. Ize, G. Buchanan, G. P. Butland, J. Greenblatt, A. Emili, and T. Palmer
Conserved Network of Proteins Essential for Bacterial Viability
J. Bacteriol., August 1, 2009; 191(15): 4732 - 4749.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
S. Das, T. Kanamoto, X. Ge, P. Xu, T. Unoki, C. L. Munro, and T. Kitten
Contribution of Lipoproteins and Lipoprotein Processing to Endocarditis Virulence in Streptococcus sanguinis
J. Bacteriol., July 1, 2009; 191(13): 4166 - 4179.
[Abstract] [Full Text] [PDF]

Home page
 Acta Biochim Biophys SinHome page
F. Shi, Y. Li, Y. Li, and X. Wang
Molecular properties, functions, and potential applications of NAD kinases
Acta Biochim Biophys Sin, May 1, 2009; 41(5): 352 - 361.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
D. A. Rodionov, P. Hebbeln, A. Eudes, J. ter Beek, I. A. Rodionova, G. B. Erkens, D. J. Slotboom, M. S. Gelfand, A. L. Osterman, A. D. Hanson, et al.
A Novel Class of Modular Transporters for Vitamins in Prokaryotes
J. Bacteriol., January 1, 2009; 191(1): 42 - 51.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
R. Zhang and Y. Lin
DEG 5.0, a database of essential genes in both prokaryotes and eukaryotes
Nucleic Acids Res., January 1, 2009; 37(suppl_1): D455 - D458.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
H. S. Seo, R. T. Cartee, D. G. Pritchard, and M. H. Nahm
A New Model of Pneumococcal Lipoteichoic Acid Structure Resolves Biochemical, Biosynthetic, and Serologic Inconsistencies of the Current Model
J. Bacteriol., April 1, 2008; 190(7): 2379 - 2387.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Poncet-Montange, L. Assairi, S. Arold, S. Pochet, and G. Labesse
NAD Kinases Use Substrate-assisted Catalysis for Specific Recognition of NAD
J. Biol. Chem., November 23, 2007; 282(47): 33925 - 33934.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
J. J. E. Bijlsma, P. Burghout, T. G. Kloosterman, H. J. Bootsma, A. de Jong, P. W. M. Hermans, and O. P. Kuipers
Development of Genomic Array Footprinting for Identification of Conditionally Essential Genes in Streptococcus pneumoniae
Appl. Envir. Microbiol., March 1, 2007; 73(5): 1514 - 1524.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
P. Hebbeln, D. A. Rodionov, A. Alfandega, and T. Eitinger
Biotin uptake in prokaryotes by solute transporters with an optional ATP-binding cassette-containing module
PNAS, February 20, 2007; 104(8): 2909 - 2914.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
A. R. Joyce, J. L. Reed, A. White, R. Edwards, A. Osterman, T. Baba, H. Mori, S. A. Lesely, B. O. Palsson, and S. Agarwalla
Experimental and Computational Assessment of Conditionally Essential Genes in Escherichia coli
J. Bacteriol., December 1, 2006; 188(23): 8259 - 8271.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. A. Ruiz-Maso, S. P. Anand, M. Espinosa, S. A. Khan, and G. del Solar
Genetic and Biochemical Characterization of the Streptococcus pneumoniae PcrA Helicase and Its Role in Plasmid Rolling Circle Replication
J. Bacteriol., November 1, 2006; 188(21): 7416 - 7425.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
A. Hunt, J. P. Rawlins, H. B. Thomaides, and J. Errington
Functional analysis of 11 putative essential genes in Bacillus subtilis.
Microbiology, October 1, 2006; 152(Pt 10): 2895 - 2907.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
M. D. Cummings, M. A. Farnum, and M. I. Nelen
Universal Screening Methods and Applications of ThermoFluor(R)
J Biomol Screen, October 1, 2006; 11(7): 854 - 863.
[Abstract] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
S. T. Cardona, C. L. Mueller, and M. A. Valvano
Identification of Essential Operons with a Rhamnose-Inducible Promoter in Burkholderia cenocepacia
Appl. Envir. Microbiol., April 1, 2006; 72(4): 2547 - 2555.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
H. Sanchez, D. Kidane, P. Reed, F. A. Curtis, M. C. Cozar, P. L. Graumann, G. J. Sharples, and J. C. Alonso
The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis
Genetics, November 1, 2005; 171(3): 873 - 883.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. E. Carver, B. Bordeau, M. D. Cummings, E. C. Petrella, M. J. Pucci, L. E. Zawadzke, B. A. Dougherty, J. A. Tredup, J. W. Bryson, J. Yanchunas Jr., et al.
Decrypting the Biochemical Function of an Essential Gene from Streptococcus pneumoniae Using ThermoFluor(R) Technology
J. Biol. Chem., March 25, 2005; 280(12): 11704 - 11712.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
W. Haas, J. Sublett, D. Kaushal, and E. I. Tuomanen
Revising the Role of the Pneumococcal vex-vncRS Locus in Vancomycin Tolerance
J. Bacteriol., December 15, 2004; 186(24): 8463 - 8471.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Y. Galperin and E. V. Koonin
'Conserved hypothetical' proteins: prioritization of targets for experimental study
Nucleic Acids Res., October 12, 2004; 32(18): 5452 - 5463.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Bouhss, M. Crouvoisier, D. Blanot, and D. Mengin-Lecreulx
Purification and Characterization of the Bacterial MraY Translocase Catalyzing the First Membrane Step of Peptidoglycan Biosynthesis
J. Biol. Chem., July 16, 2004; 279(29): 29974 - 29980.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
K. O'Dwyer, J. M. Watts, S. Biswas, J. Ambrad, M. Barber, H. Brule, C. Petit, D. J. Holmes, M. Zalacain, and W. M. Holmes
Characterization of Streptococcus pneumoniae TrmD, a tRNA Methyltransferase Essential for Growth
J. Bacteriol., April 15, 2004; 186(8): 2346 - 2354.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
A. J. Lloyd, P. E. Brandish, A. M. Gilbey, and T. D. H. Bugg
Phospho-N-Acetyl-Muramyl-Pentapeptide Translocase from Escherichia coli: Catalytic Role of Conserved Aspartic Acid Residues
J. Bacteriol., March 15, 2004; 186(6): 1747 - 1757.
[Abstract] [Full Text] [PDF]

Home page
 Infect. Immun.Home page
S. Garbom, A. Forsberg, H. Wolf-Watz, and B.-M. Kihlberg
Identification of Novel Virulence-Associated Genes via Genome Analysis of Hypothetical Genes
Infect. Immun., March 1, 2004; 72(3): 1333 - 1340.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
S. Y. Gerdes, M. D. Scholle, J. W. Campbell, G. Balazsi, E. Ravasz, M. D. Daugherty, A. L. Somera, N. C. Kyrpides, I. Anderson, M. S. Gelfand, et al.
Experimental Determination and System Level Analysis of Essential Genes in Escherichia coli MG1655
J. Bacteriol., October 1, 2003; 185(19): 5673 - 5684.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
G. T. Robertson, W.-L. Ng, R. Gilmour, and M. E. Winkler
Essentiality of clpX, but Not clpP, clpL, clpC, or clpE, in Streptococcus pneumoniae R6
J. Bacteriol., May 1, 2003; 185(9): 2961 - 2966.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.