We have constructed a series of novel eukaryotic expression systems that allows expression of transfected genes in Crithidia and Leishmania. These cells can be readily transfected with plasmid DNA by electroporation and transformants selected with various antibiotic resistances. Genes can be readily inserted using a multiple cloning site and expressed as a fusion protein with a poly-histidine tag or fused to green, blue, yellow, cyan and red fluorescent proteins at either the N or C-terminus.
Biologically active proteins can be expressed in C. fasciculata, L. amazonensis, L. mexicana or L. major promastigotes at relatively high levels and purified by affinity chromatography using a metal chelating column.
