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PMID: 9671781 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Systematic identification of essential genes by in vitro mariner mutagenesis.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 15 ·1998-07-21 ·Pages 8927-32

Akerley BJ, Rubin EJ, Camilli A, Lampe DJ, Robertson HM, Mekalanos JJ

Abstract

Although the complete DNA sequences of several microbial genomes are now available, nearly 40% of the putative genes lack identifiable functions. Comprehensive screens and selections for identifying functional classes of genes are needed to convert sequence data into meaningful biological information. One particularly significant group of bacterial genes consists of those that are essential for growth or viability. Here, we describe a simple system for performing transposon mutagenesis on naturally transformable organisms along with a technique to rapidly identify essential or conditionally essential DNA segments. We show the general utility of this approach by applying it to two human pathogens, Haemophilus influenzae and Streptococcus pneumoniae, in which we detected known essential genes and assigned essentiality to several ORFs of unknown function.

MeSH Terms
Base Sequence DNA Primers DNA Transposable Elements DNA-Binding Proteins Genes, Bacterial Haemophilus influenzae/genetics Mutagenesis Open Reading Frames Streptococcus pneumoniae/genetics Transposases
Chemicals
DNA Primers DNA Transposable Elements DNA-Binding Proteins mariner transposases Transposases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Akerley B J
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
Rubin E J
Camilli A
Lampe D J
Robertson H M
Mekalanos J J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.15.8927
Published
1998-07-21
Pages
8927-32
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC21179
Subset
IM
Grants
NIAID NIH HHS · R01 AI026289 · United States
NIAID NIH HHS · AI01237 · United States
NIAID NIH HHS · AI26289 · United States
External Links
PubMed source DOI: 10.1073/pnas.95.15.8927 PMC Full Text View journal details
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