Document Detail

Constructing Unmarked Gene Deletions in Legionella pneumophila.
MedLine Citation:
PMID:  23150396     Owner:  NLM     Status:  Publisher    
The ability to construct recombinant alleles efficiently in strains of interest, particularly unmarked deletions that reduce the potential for polar effects, is essential to studies of both pathogenesis and basic bacterial physiology. Here we describe a three-phase approach for generating unmarked deletions in Legionella pneumophila by constructing a mutant allele in E. coli using λ-Red recombination, so-called recombineering; transferring the allele onto the L. pneumophila chromosome by natural transformation; and then removing the selectable marker by utilizing the Flp site-specific recombinase. This strategy can decrease the amount of clone screening required while also increasing the percentage of the time the desired allele is obtained on the first attempt. The approach is particularly suited for constructing multiple unmarked deletions in a single strain in fewer steps than traditional methods.
Andrew Bryan; Zachary D Abbott; Michele S Swanson
Related Documents :
22817636 - Genetic modifiers of comatose mutations in drosophila: insights into neuronal nsf (n-et...
21984976 - High frequency of pik3r1 and pik3r2 mutations in endometrial cancer elucidates a novel ...
23329866 - Semiparametric inference on the penetrances of rare genetic mutations based on a case-f...
2138836 - Comparative biochemical and genetic studies of the gox-1 (hao-1) and the xdh-1 polymorp...
7858176 - Genetics and biochemistry of creutzfeldt-jakob disease in libyan jews.
1301916 - Characterization of mutations in phenotypic variants of hypoxanthine phosphoribosyltran...
Publication Detail:
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  954     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  197-212     Citation Subset:  -    
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Induction of Competence for Natural Transformation in Legionella pneumophila and Exploitation for Mu...
Next Document:  Natural Biofilm Formation with Legionella pneumophila.