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

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

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

Biology

Research output: Contribution to journal › Article (journal) › peer-review

41 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 language	English
Pages (from-to)	515-522
Journal	Antonie van Leeuwenhoek
Volume	99
Issue number	3
DOIs	https://doi.org/doi:10.1007/s10482-010-9518-1
Publication status	Published - 2011

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doi:10.1007/s10482-010-9518-1

http://dx.doi.org/10.1007/s10482-010-9518-1

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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.",

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year = "2011",

doi = "doi:10.1007/s10482-010-9518-1",

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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

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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 (journal)

SN - 0003-6072

VL - 99

SP - 515

EP - 522

JO - Antonie van Leeuwenhoek

JF - Antonie van Leeuwenhoek

IS - 3

ER -

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

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