Genome Research

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

Published online before print April 12, 2004, 10.1101/gr.1710304
Genome Res. 14:893-900, 2004
©2004 by Cold Spring Harbor Laboratory Press; ISSN 1088-9051/04 $5.00
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1710304v1
14/5/893    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Bishop, A.
Right arrow Articles by Herron, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bishop, A.
Right arrow Articles by Herron, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Methods

Systematic Insertional Mutagenesis of a Streptomycete Genome: A Link Between Osmoadaptation and Antibiotic Production

Amy Bishop, Sue Fielding, Paul Dyson1 and Paul Herron

School of Biological Sciences, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK

The model organism Streptomyces coelicolor represents a genus that produces a vast range of bioactive secondary metabolites. We describe a versatile procedure for systematic and comprehensive mutagenesis of the S. coelicolor genome. The high-throughput process relies on in vitro transposon mutagenesis of an ordered cosmid library; mutagenized cosmids with fully characterized insertions are then transferred by intergeneric conjugation into Streptomyces, where gene replacement is selected. The procedure can yield insertions in upward of 90% of genes, and its application to the entire genome is underway. The methodology could be applied to many other organisms that can receive DNA via RK2/RP4-mediated intergeneric conjugation. The system permits introduction of mutations into different genetic backgrounds and qualitative measurement of the expression of disrupted genes as demonstrated in the analysis of a hybrid histidine kinase and response regulator gene pair, osaAB, involved in osmoadaptation in Streptomyces. The independently transcribed response regulator gene, osaB, is essential for osmoadaptation; when grown with supplementary osmolyte, an osaB mutant cannot erect aerial hyphae and produces up to fivefold greater antibiotic yields than the wild-type strain.

[The sequence data from this study have been submitted to EMBL under accession nos. AJ565873 and AJ566337.]

Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.1710304. Article published online before print in April 2004.

1 Corresponding author.
E-MAIL p.j.dyson{at}swansea.ac.uk; FAX 44-1792-295447.


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


This article has been cited by other articles:


Home page
 J. Bacteriol.Home page
K. Vrancken, L. Van Mellaert, and J. Anne
Characterization of the Streptomyces lividans PspA Response
J. Bacteriol., May 15, 2008; 190(10): 3475 - 3481.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
S. Colson, G. P. van Wezel, M. Craig, E. E. E. Noens, H. Nothaft, A. M. Mommaas, F. Titgemeyer, B. Joris, and S. Rigali
The chitobiose-binding protein, DasA, acts as a link between chitin utilization and morphogenesis in Streptomyces coelicolor
Microbiology, February 1, 2008; 154(2): 373 - 382.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. A. Bennett, R. M. Aimino, and J. R. McCormick
Streptomyces coelicolor Genes ftsL and divIC Play a Role in Cell Division but Are Dispensable for Colony Formation
J. Bacteriol., December 15, 2007; 189(24): 8982 - 8992.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
L. Wang, Y. Yu, X. He, X. Zhou, Z. Deng, K. F. Chater, and M. Tao
Role of an FtsK-Like Protein in Genetic Stability in Streptomyces coelicolor A3(2)
J. Bacteriol., March 15, 2007; 189(6): 2310 - 2318.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
H. Petkovic, J. Cullum, D. Hranueli, I. S. Hunter, N. Peric-Concha, J. Pigac, A. Thamchaipenet, D. Vujaklija, and P. F. Long
Genetics of Streptomyces rimosus, the Oxytetracycline Producer
Microbiol. Mol. Biol. Rev., September 1, 2006; 70(3): 704 - 728.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
M. Y. Galperin
Structural classification of bacterial response regulators: diversity of output domains and domain combinations.
J. Bacteriol., June 1, 2006; 188(12): 4169 - 4182.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
G. Khodakaramian, S. Lissenden, B. Gust, L. Moir, P. A. Hoskisson, K. F. Chater, and M. C. M. Smith
Expression of Cre recombinase during transient phage infection permits efficient marker removal in Streptomyces
Nucleic Acids Res., February 9, 2006; 34(3): e20 - e20.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
M. I. Hutchings, P. A. Hoskisson, G. Chandra, and M. J. Buttner
Sensing and responding to diverse extracellular signals? Analysis of the sensor kinases and response regulators of Streptomyces coelicolor A3(2)
Microbiology, September 1, 2004; 150(9): 2795 - 2806.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
P. R. Herron, G. Hughes, G. Chandra, S. Fielding, and P. J. Dyson
Transposon Express, a software application to report the identity of insertions obtained by comprehensive transposon mutagenesis of sequenced genomes: analysis of the preference for in vitro Tn5 transposition into GC-rich DNA
Nucleic Acids Res., August 12, 2004; 32(14): e113 - e113.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Genes Dev. Learn. Mem.
Protein Science RNA Genome Res.
Copyright © 2004 by Cold Spring Harbor Laboratory Press.