Reverse transcription-polymerase chain reaction construction of plasmid-based, full-length cDNA libraries from Leishmania infantum for in vitro expression screening
Mem. Inst. Oswaldo Cruz
;
98(4): 477-480, June 2003. ilus, tab
Artigo
em Inglês
| LILACS
| ID: lil-344238
RESUMO
We describe a streamlined reverse transcription-polymerase chain reaction methodology for constructing full-length cDNA libraries of trypanosomatids on the basis of conserved sequences located at the 5' and 3'ends of trans-spliced mRNAs. The amplified cDNA corresponded to full-length messengers and was amenable to in vitro expression. Fractionated libraries could be rapidly constructed in a plasmid vector by the TA cloning method (Invitrogen). We believe this is useful when there are concerns over the use of restriction enzymes and phage technology as well as in cases where expression of proteins in their native conformation is desired
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Biblioteca Gênica
/
DNA de Protozoário
/
RNA de Protozoário
/
Leishmania infantum
Tipo de estudo:
Estudo diagnóstico
/
Estudo de rastreamento
Limite:
Animais
Idioma:
Inglês
Revista:
Mem. Inst. Oswaldo Cruz
Assunto da revista:
Medicina Tropical
/
Parasitologia
Ano de publicação:
2003
Tipo de documento:
Artigo
País de afiliação:
Bélgica
Instituição/País de afiliação:
Free University of Brussels/BE
/
Prince Leopold Institute of Tropical Medicine/BE
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