Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid

Abstract

Background and purpose: Toxoplasma gondii (T. gondii) is an obligatory intracellular protozoan parasite. Toxoplasmosis is highly prevalent and has a great effect on public health, therefor, there is a need for vaccine and sensitive diagnostic procedures. This study aimed at cloning and investigating the expression of ROP13 gene of T. gondii. Materials and methods: In this study, the gene was cloned in pTG19-T vector and transferred to a TOP10 strain of E.coli following ROP13 gene amplification using PCR. Then the ROP13 gene was sub cloned in expression plasmid pcDNA3. Also, pcROP13 was transferred to CHO cells and the expression level was evaluated by IFA method. Results: Cloning and sub cloning of ROP 13 gene were confirmed by PCR, sequencing and enzymatic digestion. The gene sequencing showed complete homology with a recorded sequence in the gene bank. Moreover, the expression of ROP13 gene in CHO eukaryotic cells was confirmed by IFA method. Conclusion: The results showed that ROP13 gene was successfully sub cloned into the pcDNA3 expression vector and expressed in CHO cells. Therefore, it can be used in the development of vaccines and in diagnostic tests.

Keywords

• toxoplasmosis
• rhoptry 13
• pcdna3
• gene expression