A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2)

L. T. Fernández-Martínez, R. Del Sol, M. C. Evans, S. Fielding, P. R. Herron, G. Chandra, P. J. Dyson

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB (http://​strepdb.​streptomyces.​org.​uk/​). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.
Original languageEnglish
Pages (from-to)515-522
JournalAntonie van Leeuwenhoek
Volume99
Issue number3
DOIs
Publication statusPublished - 2011

Fingerprint

Streptomyces coelicolor
Cosmids
Gene Knockout Techniques
Gene Library
Mutagenesis
Streptomyces
Open Reading Frames
Genes
Libraries
Genome
Databases

Cite this

Fernández-Martínez, L. T. ; Del Sol, R. ; Evans, M. C. ; Fielding, S. ; Herron, P. R. ; Chandra, G. ; Dyson, P. J. / A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2). In: Antonie van Leeuwenhoek. 2011 ; Vol. 99, No. 3. pp. 515-522.
@article{5141070749ef42e785abcf7d7ea4bf2c,
title = "A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2)",
abstract = "A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83{\%} of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB (http://​strepdb.​streptomyces.​org.​uk/​). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.",
author = "Fern{\'a}ndez-Mart{\'i}nez, {L. T.} and {Del Sol}, R. and Evans, {M. C.} and S. Fielding and Herron, {P. R.} and G. Chandra and Dyson, {P. J.}",
year = "2011",
doi = "doi:10.1007/s10482-010-9518-1",
language = "English",
volume = "99",
pages = "515--522",
journal = "Antonie van Leeuwenhoek",
issn = "0003-6072",
publisher = "Springer Netherlands",
number = "3",

}

A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2). / Fernández-Martínez, L. T.; Del Sol, R.; Evans, M. C.; Fielding, S.; Herron, P. R.; Chandra, G.; Dyson, P. J.

In: Antonie van Leeuwenhoek, Vol. 99, No. 3, 2011, p. 515-522.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2)

AU - Fernández-Martínez, L. T.

AU - Del Sol, R.

AU - Evans, M. C.

AU - Fielding, S.

AU - Herron, P. R.

AU - Chandra, G.

AU - Dyson, P. J.

PY - 2011

Y1 - 2011

N2 - A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB (http://​strepdb.​streptomyces.​org.​uk/​). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.

AB - A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB (http://​strepdb.​streptomyces.​org.​uk/​). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.

U2 - doi:10.1007/s10482-010-9518-1

DO - doi:10.1007/s10482-010-9518-1

M3 - Article

VL - 99

SP - 515

EP - 522

JO - Antonie van Leeuwenhoek

JF - Antonie van Leeuwenhoek

SN - 0003-6072

IS - 3

ER -