Cloning dictyostelium Paracaspase protein in the Acanthamoeba expression plasmid

Abstract EN

Acanthamoeba castellaniiand the slime mold Dictyostelium discoideumare members of Amoebozoa.

Both microorganisms were applied as model organisms in different biological studies.

A.

castellanii metacaspase (Acmcp) and D.

discoideum paracaspase (Ddpcp) proteins have been recently discovered.

Determining the function of Acmcp and Ddpcp providesavaluable information about their role in more complex organisms.This studyaimsto clonethe Ddpcpgene in the Acanthamoebaexpression plasmids.

TheDdpcpgenewas inserted in aplasmids containing the TBP Promoter Binding Factor (TPBF) gene promoters from A.castellaniiand enhanced green fluorescent protein (EGFP) as the reporter gene.

The promoters for AcanthamoebaTPBF gene was used to drive constitutive expression of EGFPproteinin stably transfected Acanthamoeba.Recombinant TPBF is able to bind DNA and activate transcription as the natural AcanthamoebaTPBF.

The results showed a sucssesful constract of production of recombinant Ddpcpgene in pTPBfplasmid thatwill be able touse later to provide a valuable information about the function of Ddpcp in the Acanthamoeba parasite.