Trypanosoma cruzi: cloning and characterization of two genes whose expression is up-regulated in metacyclic trypomastigotes
| Autor: | Stenio Perdigão Fragoso, Sueli Fumie Yamada-Ogatta, Bruno Dallagiovana Muniz, Samuel Goldenberg, Andréa Rodrigues Ávila, Viviane Monteiro-Góes, Helena Keiko Toma, Celso Vataru Nakamura, Marco Aurélio Krieger, Maria Cristina M. Motta |
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| Rok vydání: | 2004 |
| Předmět: |
Life Cycle Stages
medicine.diagnostic_test biology Cloning Organism Trypanosoma cruzi Veterinary (miscellaneous) Genes Protozoan Nucleic acid sequence Gene Expression biology.organism_classification Molecular biology Up-Regulation Blot Infectious Diseases Western blot Insect Science Kinetoplast Gene expression medicine Animals Parasitology Gene Southern blot |
| Zdroj: | Acta Tropica. 90:171-179 |
| ISSN: | 0001-706X |
| DOI: | 10.1016/j.actatropica.2003.10.018 |
| Popis: | The differentiation of epimastigotes into metacyclic trypomastigotes (metacyclogenesis) involves the transformation of a replicative non-infective form of Trypanosoma cruzi into a non-replicative infective stage. The study of genes with stage-specific expression may provide insight into the mechanisms involved in the regulation of gene expression in this parasite. We cloned and characterized two genes whose expression is up-regulated in metacyclic trypomastigote, those encoding metacyclin-II (Met-II) and metacyclin-III (Met-III). Nucleotide sequence analysis identified no sequence similarity with sequences available from genetic databases. The deduced amino acid sequences of the genes indicated that Met-III encodes a basic polypeptide whereas Met-II encodes an acidic polypeptide. Northern and Western blot analyses showed that Met-II and Met-III were expressed by metacyclic trypomastigotes, but not by epimastigotes. Antisera directed against the recombinant Met-II and Met-III proteins recognized two polypeptides on Western blots: a 16-kDa and a 24-kDa polypeptide. Immunocytochemistry analysis using electron microscopy showed that metacyclin-II is localized mainly at the kinetoplast whereas metacyclin-III is localized at the nucleus of the parasite. Southern blot analysis, using genomic DNA and T. cruzi chromosomes separated by pulsed-field gel electrophoresis, indicated that these genes were present as single copies on different chromosomes of T. cruzi Dm28c. |
| Databáze: | OpenAIRE |
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