SHORT COMMUNICATION - Reverse Transcription-Polymerase Chain Reaction Construction of Plasmid-based, Full-length cDNA Libraries from Leishmania infantum for in Vitro Expression Screening

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 messe...

Full description

Saved in:
Bibliographic Details
Published in:Memórias do Instituto Oswaldo Cruz 2003-07, Vol.98 (4)
Main Authors: Bernard Couvreur, Alex Bollen, Dominique Le Ray, Jean-Claude Dujardin
Format: Article
Language:English
Subjects:
kinetoplastid - cDNA libraries - spliced leader
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1678-8060