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Colony polymerase chain reaction of stably transfected Trypanosoma cruzi grown on solid medium
Santos, Wagner G. dos; Metcheva, Ivelina; Buck, Gregory A.
Afiliação
  • Santos, Wagner G. dos; Virginia Commonwealth University. Medical College of Virginia Campus. Department of Microbiology and Immunology.
  • Metcheva, Ivelina; Virginia Commonwealth University. Medical College of Virginia Campus. Department of Microbiology and Immunology.
  • Buck, Gregory A; Virginia Commonwealth University. Medical College of Virginia Campus. Department of Microbiology and Immunology.
Mem. Inst. Oswaldo Cruz ; 95(1): 111-4, Jan.-Feb. 2000. ilus
Article em En | LILACS | ID: lil-251322
Biblioteca responsável: BR1.1
RESUMO
Tools for the genetic manipulation of Trypanosoma cruzi are largely unavailable, although several vectors for transfection of epimastigotes and expression of foreign or recombinant genes have been developed. We have previously constructed several plasmid vectors in which recombinant genes are expressed in T. cruzi using the rRNA promoter. In this report, we demonstrate that one of these vectors can simultaneously mediate expression of neomycin phosphotransferase and green fluorescent protein when used to stably transfect cultured epimastigotes. These stably transfected epimastigotes can be selected and cloned as unique colonies on solid medium. We describe a simple colony PCR approach to the screening of these T. cruzi colonies for relevant genes. Thus, the methodologies outlined herein provide important new tools for the genetic dissection of this important parasite.
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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Trypanosoma cruzi / Transfecção / Reação em Cadeia da Polimerase Limite: Animals Idioma: En Revista: Mem. Inst. Oswaldo Cruz Assunto da revista: MEDICINA TROPICAL / PARASITOLOGIA Ano de publicação: 2000 Tipo de documento: Article País de publicação: Brasil
Texto completo
Adicionar na Minha BVS
Imprimir
XML
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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Trypanosoma cruzi / Transfecção / Reação em Cadeia da Polimerase Limite: Animals Idioma: En Revista: Mem. Inst. Oswaldo Cruz Assunto da revista: MEDICINA TROPICAL / PARASITOLOGIA Ano de publicação: 2000 Tipo de documento: Article País de publicação: Brasil