Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid

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Alizadeh Paria; Daryani Ahamd; AHMADPOUR EHSAN; KAZEMI TOHID; SPOTIN ADEL; Mahami Oskoui Mahmoud; Azadi Yaghob; Rajabi Saba; SHANEHBANDI DARIUSH

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Journal: JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES Year:2019 | Volume:28 | Issue:169 Page(s): 26-35

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Title

Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid

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Keywords

toxoplasmosisQ3

Rhoptry 13Q1

pcDNA3Q1

gene expressionQ1

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.

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APA: Copy

Alizadeh, Paria, Daryani, Ahamd, AHMADPOUR, EHSAN, KAZEMI, TOHID, SPOTIN, ADEL, Mahami Oskoui, Mahmoud, Azadi, Yaghob, Rajabi, Saba, & SHANEHBANDI, DARIUSH. (2019). Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid. JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, 28(169 ), 26-35. SID. https://sid.ir/paper/46101/en

Vancouver: Copy

Alizadeh Paria, Daryani Ahamd, AHMADPOUR EHSAN, KAZEMI TOHID, SPOTIN ADEL, Mahami Oskoui Mahmoud, Azadi Yaghob, Rajabi Saba, SHANEHBANDI DARIUSH. Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid. JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES[Internet]. 2019;28(169 ):26-35. Available from: https://sid.ir/paper/46101/en

IEEE: Copy

Paria Alizadeh, Ahamd Daryani, EHSAN AHMADPOUR, TOHID KAZEMI, ADEL SPOTIN, Mahmoud Mahami Oskoui, Yaghob Azadi, Saba Rajabi, and DARIUSH SHANEHBANDI, “Cloning and Expression of Toxoplasma gondii Rhoptry13 Gene in pcDNA3 Plasmid,” JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, vol. 28, no. 169 , pp. 26–35, 2019, [Online]. Available: https://sid.ir/paper/46101/en

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