Specificity of modified Drosophila mariner transposons in the identification of Leishmania genes.
Genetic manipulation of the protozoan Leishmania has led to a better understanding of the survival and development of these pathogens within their hosts. The association of the Leishmania genome sequencing information with the ability of transposons to introduce or destroy phenotypes allows a global perspective on the role and importance of genes in cellular pathways. Herein we report the construction and testing of mariner transposable elements carrying the neomycin phosphotransferase, green fluorescent protein, or beta-glucuronidase genes as reporters for translational fusion events. We demonstrate that the expression of the reporter genes will occur only when the genes are inserted in-frame within predicted genes. Our results not only add to the mariner toolkit for gene manipulation but also strengthen the evidence that the mariner system is a reliable means for the study of gene expression in Leishmania.
Augusto
Marlei J
MJ
Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo. Av. Bandeirantes 3900, 14049-900 Ribeirão Preto-SP, Brazil.
Squina
Fabio M
FM
Marchini
Julio F M
JF
Dias
Fabricio C
FC
Tosi
Luiz R O
LR
D000818:Animals; D004251:DNA Transposable Elements / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics; D019868:Kanamycin Kinase / Q000235:genetics; D007891:Leishmania / Q000235:genetics*; D016254:Mutagenesis, Insertional; D013045:Species Specificity; D014162:Transfection; D019895:Transposases
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C111652:mariner transposases; D049452:Green Fluorescent Proteins; D019868:Kanamycin Kinase; D019895:Transposases