allele replacement
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2018 ◽  
Vol 74 (3) ◽  
pp. 618-624 ◽  
Author(s):  
Isabelle Accoceberry ◽  
Célia Couzigou ◽  
Valérie Fitton-Ouhabi ◽  
Nicolas Biteau ◽  
Thierry Noël

Fly ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. 53-64 ◽  
Author(s):  
Abigail M. Lamb ◽  
Elizabeth A. Walker ◽  
Patricia J. Wittkopp
Keyword(s):  

Genetics ◽  
2014 ◽  
Vol 198 (3) ◽  
pp. 859-866 ◽  
Author(s):  
William G. Alexander ◽  
Drew T. Doering ◽  
Chris Todd Hittinger

2013 ◽  
Vol 60 (2) ◽  
pp. 109-119 ◽  
Author(s):  
Jun Gao ◽  
Fengling Kan ◽  
Jacy L. Wagnon ◽  
Aaron J. Storey ◽  
Reine U. Protacio ◽  
...  

2011 ◽  
Vol 77 (10) ◽  
pp. 3532-3535 ◽  
Author(s):  
Caray A. Walker ◽  
Willie Donachie ◽  
David G. E. Smith ◽  
Michael C. Fontaine

ABSTRACTA two-step allele replacement mutagenesis procedure, using a conditionally replicating plasmid, was developed to allow the creation of targeted, marker-free mutations inCorynebacterium pseudotuberculosis. The relationship between homologous sequence length and recombination frequency was determined, and enhanced plasmid excision was observed due to the rolling-circle replication of the mutagenesis vector. Furthermore, an antibiotic enrichment procedure was applied to improve the recovery of mutants. Subsequently, as proof of concept, a marker-free,cp40-deficient mutant ofC. pseudotuberculosiswas constructed.


2009 ◽  
Vol 75 (20) ◽  
pp. 6496-6503 ◽  
Author(s):  
Carolina M. López ◽  
Drew A. Rholl ◽  
Lily A. Trunck ◽  
Herbert P. Schweizer

ABSTRACT Burkholderia pseudomallei is the etiologic agent of melioidosis, a rare but serious tropical disease. In the United States, genetic research with this select agent bacterium is strictly regulated. Although several select agent compliant methods have been developed for allelic replacement, all of them suffer from some drawbacks, such as a need for specific host backgrounds or use of minimal media. Here we describe a versatile select agent compliant allele replacement system for B. pseudomallei based on a mobilizable vector, pEXKm5, which contains (i) a multiple cloning site within a lacZα gene for facile cloning of recombinant DNA fragments, (ii) a constitutively expressed gusA indicator gene for visual detection of merodiploid formation and resolution, and (iii) elements required for resolution of merodiploids using either I-SceI homing endonuclease-stimulated recombination or sacB-based counterselection. The homing endonuclease-based allele replacement system is completed by pBADSce, which contains an araC-PBAD-I-sceI expression cassette for arabinose-inducible I-SceI expression and a temperature-sensitive pRO1600 replicon for facile plasmid curing. Complementing these systems is the improved Δasd Escherichia coli mobilizer strain RHO3. This strain is susceptible to commonly used antibiotics and allows nutritional counterselection on rich media because of its diaminopimelic acid auxotrophy. The versatility of the I-SceI- and sacB-based methods afforded by pEXKm5 in conjunction with E. coli RHO3 was demonstrated by isolation of diverse deletion mutants in several clinical, environmental, and laboratory B. pseudomallei strains. Finally, sacB-based counterselection was employed to isolate a defined chromosomal fabD(Ts) allele that causes synthesis of a temperature-sensitive FabD, an essential fatty acid biosynthesis enzyme.


2006 ◽  
Vol 27 (2) ◽  
pp. 159-164 ◽  
Author(s):  
Katarzyna Dzierżanowska-Fangrat ◽  
Katarzyna Semczuk ◽  
Paulina Górska ◽  
Stefania Giedrys-Kalemba ◽  
Maria Kochman ◽  
...  

2004 ◽  
pp. 269-280 ◽  
Author(s):  
Eva-Maria Borst ◽  
György Pósfai ◽  
Frank Pogoda ◽  
Martin Messerle
Keyword(s):  

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