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Selection analyses of insertional mutants using subgenic-resolution arrays.
Badarinarayana, V; Estep, P W; Shendure, J; Edwards, J; Tavazoie, S; Lam, F; Church, G M.
Afiliación
  • Badarinarayana V; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Nat Biotechnol ; 19(11): 1060-5, 2001 Nov.
Article en En | MEDLINE | ID: mdl-11689852
We describe a method of genome-wide analysis of quantitative growth phenotypes using insertional mutagenesis and DNA microarrays. We applied the method to assess the fitness contributions of Escherichia coli gene domains under specific growth conditions. A transposon library was subjected to competitive growth selection in Luria-Bertani (LB) and in glucose minimal media. Transposon-containing genomic DNA fragments from the selected libraries were compared with the initial unselected transposon insertion library on DNA microarrays to identify insertions that affect fitness. Genes involved in the biosynthesis of nutrients not provided in the growth medium were found to be significantly enriched in the set of genes containing negatively selected insertions. The data also identify fitness contributions of several uncharacterized genes, including putative transcriptional regulators and enzymes. The applicability of this high-resolution array selection in other species is discussed.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mutagénesis Insercional / Huella de ADN / Análisis de Secuencia por Matrices de Oligonucleótidos / Escherichia coli Tipo de estudio: Evaluation_studies Idioma: En Revista: Nat Biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2001 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mutagénesis Insercional / Huella de ADN / Análisis de Secuencia por Matrices de Oligonucleótidos / Escherichia coli Tipo de estudio: Evaluation_studies Idioma: En Revista: Nat Biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2001 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos