Data/hora: 13/06/2024 21:48:56

Provedor de dados: | 19
País: | Japan
Título: | Establishment of an in vitro Transgene Expression System in Epimastigotes of　Trypanosoma congolense
Autores: | Sakurai, Tatsuya; Tanaka, Miho; Shin-ichiro, Kawazu; Inoue, Noboru; 井上, 昇.
Data: | 2009-03
Ano: | 2009
Palavras-chave: | Trypanosoma congolense; Epimastigotes; In vitro metacyclogenesis; Transfection; Transgene expression.
Resumo: | The epimastigote form (EMF) of Trypanosoma congolense appears in the late tsetse infective stage. Epimastigotes adhere to the tsetse proboscis, proliferate in this region, and differentiate into mammal-infective metacyclic forms (MCFs). This differentiation is called metacyclogenesis and is indispensable for cyclical transmission of the parasite. Although an in vitro culture method reproducing metacyclogenesis was established several decades ago, few genetic tools have been utilized to elucidate the molecular mechanisms underlying T. congolense metacyclogenesis. In this study, we established a transgene expression system in the EMF of T. congolense IL3000; the EMF was successfully cultured and observed to undergo metacyclogenesis in vitro. The newly constructed expression vector pSAK was designed for integration into the repetitive α-β tubulin locus of the T. congolense genome. pSAK/enhanced green fluorescent protein (eGFP) was transfected into the EMF and procyclic form (PCF), which were cultured in vitro by electroporation. Both EMFs and PCFs expressing eGFP were successfully generated. The eGFP expressing EMFs differentiated into MCFs that continued to express eGFP. The in vitro transgenic EMF generation method is expected to contribute to the elucidation of molecular mechanisms underlying metacyclogenesis.
Idioma: | Inglês
Identificador: | http://ir.obihiro.ac.jp/dspace/handle/10322/2677
Editor: | Elsevier Ireland Ltd.
Formato: | application/pdf
Fonte: | Elsevier Ireland Ltd.