Date of Award

2009

Degree Type

Thesis

Degree Name

M.Sc.

Department

Chemistry and Biochemistry

First Advisor

Ananvoranich, Sirinart (Chemistry and Biochemistry)

Keywords

Biochemistry.

Rights

CC BY-NC-ND 4.0

Abstract

Ribonuclease activity within the RNAi mechanism has been thoroughly described in model organisms such as Caenorhabditis elegans. However, RNAi is still largely a mystery in Apicomplexan parasites. Our group previously has reported that specific gene silencing via the introduction of dsRNA in Toxoplasma gondii can be achieved. Furthermore, a key protein (Argonaute) has been characterized. In this study, through the use of bioinformatic tools, several putative proteins that may act upon the RNAi mechanism have been identified. Furthermore in vitro dsRNA-directed ribonuclease activity and dsRNA-specific ribonuclease activity in Toxoplasma gondii lysates were assessed. Lastly, using a His construct, the Argonaute protein in T. gondii was successfully isolated from inclusion bodies and characterized in terms of its interaction with RNA species. This study demonstrates the presence of an RNAi-like mechanism in T. gondii and can lead to determination of future drug targets.

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