ArticleMolecular and biochemical parasitology2011
A rapid, efficient and economical method for generating leishmanial gene targeting constructs.
Article in Molecular and biochemical parasitology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- A high-throughput protein tagging toolkit that retains endogenous untranslated regions for studying gene regulation in kinetoplastids.Open biology · 2025Article
- Article
- A cytoskeletal protein complex is essential for division of intracellular amastigotes ofThe Journal of biological chemistry · 2020Article
- Purine-responsive expression of theThe Journal of biological chemistry · 2020Article
- PRMT7 regulates RNA-binding capacity and protein stability in Leishmania parasites.Nucleic acids research · 2020Article
- Ascorbate-Dependent Peroxidase (APX) from Leishmania amazonensis Is a Reactive Oxygen Species-Induced Essential Enzyme That Regulates Virulence.Infection and immunity · 2019Article
- Leishmania donovani Lipophosphoglycan Increases Macrophage-Dependent Chemotaxis of CXCR6-Expressing Cells via CXCL16 Induction.Infection and immunity · 2019Article
- A MFS-like plasma membrane transporter required for Leishmania virulence protects the parasites from iron toxicity.PLoS pathogens · 2018Article
- The iron-dependent mitochondrial superoxide dismutase SODA promotesThe Journal of biological chemistry · 2017Article
- GMP reductase and genetic uncoupling of adenylate and guanylate metabolism in Leishmania donovani parasites.Molecular and biochemical parasitology · 2016Article
- A Trypanosomatid Iron Transporter that Regulates Mitochondrial Function Is Required for Leishmania amazonensis Virulence.PLoS pathogens · 2016Article
- Integrating ribosomal promoter vectors that offer a choice of constitutive expression profiles in Leishmania donovani.Molecular and biochemical parasitology · 2015Article
- Regulation and biological function of a flagellar glucose transporter in Leishmania mexicana: a potential glucose sensor.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2015Article
- A dual luciferase system for analysis of post-transcriptional regulation of gene expression in Leishmania.Molecular and biochemical parasitology · 2014Article
- Metabolic reprogramming during purine stress in the protozoan pathogen Leishmania donovani.PLoS pathogens · 2014Article
- Genetically modified organisms and visceral leishmaniasis.Frontiers in immunology · 2014Review
- KHARON1 mediates flagellar targeting of a glucose transporter in Leishmania mexicana and is critical for viability of infectious intracellular amastigotes.The Journal of biological chemistry · 2013Article
- Adenine and adenosine salvage in Leishmania donovani.Molecular and biochemical parasitology · 2013Article
- Purine salvage in Leishmania: complex or simple by design?Trends in parasitology · 2012Review
- Genetic dissection of pyrimidine biosynthesis and salvage in Leishmania donovani.The Journal of biological chemistry · 2012Article
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Authors and funding
4 authors.
Funding
Abstract
Targeted gene replacement is a powerful tool in Leishmania genetics that can be time-consuming to implement. One tedious aspect that delays progress is the multi-step construction of gene targeting vectors. To accelerate this process, we developed a streamlined method that allows the assembly of a complete targeting vector from all its constituent parts in a single-step multi-fragment ligation. The individual components to be assembled are flanked by sites for the restriction endonuclease SfiI that generates non-identical, non-palindromic three base 3'-overhangs designed to allow annealing and ligation of the parts only in the proper order. The method was optimized by generating constructs for targeting the Leishmania donovani inosine monophosphate dehydrogenase gene (LdIMPDH) encoding six different drug resistance markers, and was found to be rapid and efficient. These constructs were successfully employed to generate heterozygous LdIMPDH gene replacement mutants. This method is adaptable for generating targeting vectors for a variety of species.
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