title: Expression of fluorescent genes in Trypanosoma cruzi and Trypanosoma rangeli (Kinetoplastida: Trypanosomatidae): its application to parasite-vector biology.
abstract: Two Trypanosoma cruzi-derived cloning vectors, pTREX-n and pBs:CalB1/CUB01, were used to drive the expression of green fluorescent protein (GFP) and DsRed in Trypanosoma rangeli Tejera, 1920, and Trypanosoma cruzi Chagas, 1909, isolates, respectively. Regardless of the species, group, or strain, parasites harboring the transfected constructs as either episomes or stable chromosomal integrations showed high-level expression of fluorescent proteins. Tagged flagellates of both species were used to experimentally infect Rhodnius prolixus Stal, 1953. In infected bugs, single or mixed infections of T. cruzi and T. rangeli displayed the typical cycle of each species, with no apparent interspecies interactions. In addition, infection of kidney monkey cells (LLC-MK2) with GFP-T. cruzi showed that the parasite retained its fluorescent tag while carrying out its life cycle within cultured cells. The use of GFP-tagged parasites as a tool for biological studies in experimental hosts is discussed, as is the application of this method for copopulation studies of same-host parasites.
authors.affiliation: Instituto de Biología Experimental, Universidad Central de Venezuela, Caracas 1041A, Venezuela. palmiragt@hotmail.com
authors.forename: Palmira
authors.initials: P
authors.lastname: Guevara
authors.forename: Manuel
authors.initials: M
authors.lastname: Dias
authors.forename: Agustina
authors.initials: A
authors.lastname: Rojas
authors.forename: Gladys
authors.initials: G
authors.lastname: Crisante
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Abreu-Blanco
authors.forename: Eufrozina
authors.initials: E
authors.lastname: Umezawa
authors.forename: Martin
authors.initials: M
authors.lastname: Vazquez
authors.forename: Mariano
authors.initials: M
authors.lastname: Levin
authors.forename: Nestor
authors.initials: N
authors.lastname: Añez
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Ramirez
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D000882:Haplorhini; D007668:Kidney; D008018:Life Cycle Stages; D008164:Luminescent Proteins / Q000235:genetics*; D012239:Rhodnius / Q000469:parasitology; D014162:Transfection; D014345:Trypanosoma / Q000235:genetics* / Q000254:growth & development; D014349:Trypanosoma cruzi / Q000235:genetics / Q000254:growth & development; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485192:green fluorescent protein, Aequorea victoria