Restriction digests of the clone give the following sizes (kb): PstI--4.8, 0.65, 0.2; BamHI--5.6; EcoRI--5.6; KpnI--5.6. The neoR gene can be used as a selectable marker in Trypanosoma cruzi and Leishmania. Increasing the concentration of G418 in the media increases copy number and expression. Shuttle expression vector for Trypanosoma cruzi and Leishmania. Transfected DNA is maintained in transformed parasites extrachromosomally as circular head-to-tail tandem repeats. DNA can be rescued from transformed parasites and propagated in E. coli after linearizing and religation to produce single circular copies. The order of the major features in this plasmid is: SacI - SacII - 5' flanking region of gGAPDH I - SpeI/MCS/XhoI - PstI - gGAPDH I/gGAPDH II intergenic region - neoR - gGAPDH II 3' flanking region - KpnI - ampR - pBluescript KS. |
