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PMID: 11689852 Published · ppublish English Comparative Study Evaluation Study Journal Article

Selection analyses of insertional mutants using subgenic-resolution arrays.

Nature biotechnology ·Vol. 19 ·No. 11 ·2001-11-00 ·Pages 1060-5

Badarinarayana V, Estep PW, Shendure J, Edwards J, Tavazoie S, Lam F, Church GM

Abstract

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.

MeSH Terms
Culture Media DNA Footprinting/methods DNA Transposable Elements Escherichia coli/genetics,growth & development Escherichia coli Proteins/genetics,physiology Genome, Bacterial Genomic Library Mutagenesis, Insertional/methods Oligonucleotide Array Sequence Analysis/methods Phenotype
Chemicals
Culture Media DNA Transposable Elements Escherichia coli Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Badarinarayana V
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Estep P W
Shendure J
Edwards J
Tavazoie S
Lam F
Church G M
Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1087-0156
Published
2001-11-00
Pages
1060-5
Language
English
Region
United States
NLM ID
9604648
Subset
IM
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