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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 98, No. 4, 2003, pp. 477-480
Bioline Code: oc03104
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 98, No. 4, 2003, pp. 477-480

 en SHORT COMMUNICATION - Reverse Transcription-Polymerase Chain Reaction Construction of Plasmid-based, Full-length cDNA Libraries from Leishmania infantum check for this species in other resources for in Vitro Expression Screening
Bernard Couvreur; Alex Bollen; Dominique Le Ray & Jean-Claude Dujardin

Abstract

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.

Keywords
kinetoplastid - cDNA libraries - spliced leader

 
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