Document Detail


One-step generation of double-allele gene replacement mutants in Leishmania donovani.
MedLine Citation:
PMID:  19028498     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Due to the apparent lack of sexual recombination in Leishmania spp., gene replacement strategies in this diploid organism necessitate the subsequent targeting of two gene alleles by using two constructs, bearing different antibiotic resistance markers. This approach is time consuming and often gives rise to spontaneous amplification of the targeted gene, nullifying efforts to create functional null mutants. Here, we show that by using two homologous recombination constructs in a co-transfection of Leishmania donovani promastigotes, we can obtain double-allele gene replacement mutants. This approach reduces the time required for the generation of functional null mutants and the number of in vitro passage cycles, potentially limiting culture-associated artefacts.
Authors:
Gabi Ommen; Sabine Lorenz; Joachim Clos
Related Documents :
6749598 - Cold-sensitive cell-division-cycle mutants of yeast: isolation, properties, and pseudor...
15141958 - Identification of botrytis cinerea genes up-regulated during infection and controlled b...
22209168 - A wd40-repeat protein controls proanthocyanidin and phytomelanin pigmentation in the se...
17456188 - Mutagenesis of the carboxy terminal protease ctpa decreases desiccation tolerance in rh...
23382218 - Divergent transcription of long noncoding rna/mrna gene pairs in embryonic stem cells.
23968928 - Molecular cloning and expression analysis of pdr1-like gene in ginseng subjected to sal...
Publication Detail:
Type:  Journal Article     Date:  2008-11-08
Journal Detail:
Title:  International journal for parasitology     Volume:  39     ISSN:  1879-0135     ISO Abbreviation:  Int. J. Parasitol.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-02-16     Completed Date:  2010-01-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0314024     Medline TA:  Int J Parasitol     Country:  England    
Other Details:
Languages:  eng     Pagination:  541-6     Citation Subset:  IM    
Affiliation:
Leishmaniasis Research Group, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht St 74, D-20359 Hamburg, Germany.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Alleles*
Animals
Blotting, Western / methods
Cells, Cultured
DNA, Protozoan / genetics
DNA, Recombinant / genetics
Drug Resistance, Microbial / genetics
Feasibility Studies
Gene Targeting / methods
Genes, Protozoan
Leishmania donovani / genetics*,  growth & development
Macrophages / parasitology
Mutagenesis, Insertional / methods
Protozoan Proteins / genetics
Reverse Transcriptase Polymerase Chain Reaction / methods
Transfection / methods*
Chemical
Reg. No./Substance:
0/DNA, Protozoan; 0/DNA, Recombinant; 0/Protozoan Proteins

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


Previous Document:  Apicomplexan cytoskeleton and motors: Key regulators in morphogenesis, cell division, transport and ...
Next Document:  Optimization of the surfaces used to capture antibodies from single hybridomas reduces the time requ...