Expression of Fluorescent Genes in Trypanosoma cruzi and Trypanosoma rangeli (Kinetoplastida: Trypanosomatidae): Its Application to Parasite-Vector Biology

• Published in: Journal of medical entomology 2005-01, Vol.42 (1), p.48-56
• Main Authors: Guevara, Palmira, Dias, Manuel, Rojas, Agustina, Crisante, Gladys, Abreu-Blanco, Maria Teresa, Umezawa, Eufrozina, Vazquez, Martin, Levin, Mariano, Añez, Nestor, Ramirez, Jose Luis
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Cell Line; cell lines; Cells, Cultured; DsRed; fluorescent labeling; Fundamental and applied biological sciences. Psychology; Gene Expression; Genetic Vectors; green fluorescent protein; Green Fluorescent Proteins - genetics; Haplorhini; host-parasite relationships; insect vectors; Kidney; Life Cycle Stages; Luminescent Proteins - genetics; Medically important nuisances and vectors, pests of stored products and materials: population survey and control; mixed infection; plasmid vectors; Red Fluorescent Protein; reporter genes; Rhodnius - parasitology; Rhodnius prolixus; Transfection; Trypanosoma - genetics; Trypanosoma - growth & development; Trypanosoma cruzi; Trypanosoma cruzi - genetics; Trypanosoma cruzi - growth & development; Trypanosoma rangeli; VECTOR/PATHOGEN/HOST INTERACTION, TRANSMISSION; Vectors. Intermediate hosts
• ISSN: 0022-2585; 1938-2928
• DOI: 10.1603/0022-2585%282005%29042%5B0048%3AEOFGIT%5D2.0.CO%3B2

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.