Stable integrative transformation of Trypanosoma brucei that occurs exclusively by homologous recombination.
A calmodulin-neomycin-resistance fusion gene was introduced into Trypanosoma brucei by electroporation, and stably transformed cell lines were obtained. In all of the transformants, the fusion gene had integrated into the host genome at the cognate locus, evidently by homologous recombination within flanking calmodulin DNA. This unusual observation distinguishes trypanosomes as the only eukaryote other than yeast known to undergo gene targeting in essentially 100% of the stable transformants. It should now be possible to systematically manipulate the trypanosome genome, directing predetermined mutations to virtually any chromosomal locus.
Eid
J
J
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205-2185.
Sollner-Webb
B
B
D000818:Animals; D002147:Calmodulin / Q000235:genetics*; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D010957:Plasmids; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection; D014170:Transformation, Genetic; D014346:Trypanosoma brucei brucei / Q000235:genetics*
D002147:Calmodulin; D004247:DNA