Recombinant protein expression in Acanthamoeba castellanii

Area of Science:

• Biotechnology and Molecular Biology
• Cell Biology and Protozoology

Background:

• The demand for improved protein production systems is ongoing.
• Novel expression hosts are continuously explored for enhanced efficiency, quality, and versatility.

Purpose of the Study:

• To investigate Acanthamoeba as a novel system for recombinant eukaryotic protein production.
• To develop an efficient transfection method for Acanthamoeba.
• To assess the capacity of Acanthamoeba castellanii for glycoprotein expression.

Main Methods:

• Plasmid vectors utilizing the TBP promoter were constructed for Acanthamoeba expression.
• Firefly luciferase reporter gene expression was used to optimize transfection protocols.
• Chikungunya virus E2 protein expression was compared across E. coli, Acanthamoeba, and mammalian cell systems.

Main Results:

• Successful expression of the firefly luciferase reporter gene was achieved.
• Acanthamoeba castellanii demonstrated expression of the Chikungunya virus E2 protein.
• The E2 protein from Acanthamoeba showed an intermediate molecular weight compared to E. coli and mammalian cells, suggesting potential glycosylation.

Conclusions:

• Acanthamoeba represents a promising protozoan host for recombinant protein production.
• The system shows potential for cost-effective and efficient eukaryotic expression.
• Further investigation into glycosylation capabilities in Acanthamoeba is warranted.