CLONING LINEAR EPITOPES OF P14-3-3 ANTIGEN INTO PEGFP-N1 VECTOR AS A DNA VACCINE MODEL TO INDUCE IMMUNO IMMUNITY AGAINST HYDATIDOSIS AND ITS EXPRESSION IN THE CHO CELL LINE

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MESRI R.; ESMAELIZAD M.; ANGAJI S.A.; TAHMASEB M.; AHMAD ZADEH M.; MOHAMMADI S.

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Journal: Armaghane Danesh Year:2015 | Volume:20 | Issue:8 (103) Page(s): 666-676

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Title

CLONING LINEAR EPITOPES OF P14-3-3 ANTIGEN INTO PEGFP-N1 VECTOR AS A DNA VACCINE MODEL TO INDUCE IMMUNO IMMUNITY AGAINST HYDATIDOSIS AND ITS EXPRESSION IN THE CHO CELL LINE

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Keywords

ECHINOCOCCUS GRANULOSUSQ3

DNA VACCINEQ2

CLONINGQ1

P14-3-3Q2

Abstract

Background & aim: Hydatidosis is a zoonotic disease caused by infection with the larvae of ECHINOCOCCUS GRANULOSUS. Different antigens produced in larval stage of this parasite that recombinant vaccine base these antigens created significant immunity in infected animals. One of the important antigens is P14-3-3 that its recombinant antigen created considerable immunity in mouse models. In the present study, according to the high immunity of antigen epitopes region, the coding sequence of T-cell epitopes of P14-3-3 was cloned into pEGFP-N1vector in order to produce an effective DNA VACCINE model to stimulate high level of Th1 immune response.Methods: In the current study, bioinformatics tools were used for the prediction of linear T-Cell epitopes of ECHINOCOCCUS GRANULOSUS P14-3-3 antigen. The nucleotide coding sequence of these epitopes was synthesized by PCR. The ampliqon was digested with XhoI restriction enzyme and cloned into pEGFP–N1 vector which had been purificated by modified by the Sambrook method with CaCl2 and PEG6000.Positive colony was selected by direct colony PCR and confirmed by the sequencing. The evaluation of its expression in Eukaryotic cells was done by transformed of CHO cell line with electroporation.Results: Linear T-cell epitopes of ECHINOCOCCUS GRANULOSUS P14-3-3 after prediction, synthesis and amplification were successfully cloned into pEGFP-N1 vector that purificated by new method with maximum vector and minimum RNA concentration. The expression of new construct in CHO cell line as a eukaryotic cells achievement by fluorescent microscope will be used as a DNA VACCINE model for the evaluation of immuno response in mouse models.Conclusion: Successfully CLONING of the linear T-cell epitppes coding sequence of ECHINOCOCCUS GRANULOSUS P14-3-3 antigen into pEGFP-N1 verified by sequencing and fluorscent microscope images demonstrated the expression of recombinant protein in CHO cell line.

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APA: Copy

MESRI, R., ESMAELIZAD, M., ANGAJI, S.A., TAHMASEB, M., AHMAD ZADEH, M., & MOHAMMADI, S.. (2015). CLONING LINEAR EPITOPES OF P14-3-3 ANTIGEN INTO PEGFP-N1 VECTOR AS A DNA VACCINE MODEL TO INDUCE IMMUNO IMMUNITY AGAINST HYDATIDOSIS AND ITS EXPRESSION IN THE CHO CELL LINE. ARMAGHAN DANESH, 20(8 (103)), 666-676. SID. https://sid.ir/paper/78179/en

Vancouver: Copy

MESRI R., ESMAELIZAD M., ANGAJI S.A., TAHMASEB M., AHMAD ZADEH M., MOHAMMADI S.. CLONING LINEAR EPITOPES OF P14-3-3 ANTIGEN INTO PEGFP-N1 VECTOR AS A DNA VACCINE MODEL TO INDUCE IMMUNO IMMUNITY AGAINST HYDATIDOSIS AND ITS EXPRESSION IN THE CHO CELL LINE. ARMAGHAN DANESH[Internet]. 2015;20(8 (103)):666-676. Available from: https://sid.ir/paper/78179/en

IEEE: Copy

R. MESRI, M. ESMAELIZAD, S.A. ANGAJI, M. TAHMASEB, M. AHMAD ZADEH, and S. MOHAMMADI, “CLONING LINEAR EPITOPES OF P14-3-3 ANTIGEN INTO PEGFP-N1 VECTOR AS A DNA VACCINE MODEL TO INDUCE IMMUNO IMMUNITY AGAINST HYDATIDOSIS AND ITS EXPRESSION IN THE CHO CELL LINE,” ARMAGHAN DANESH, vol. 20, no. 8 (103), pp. 666–676, 2015, [Online]. Available: https://sid.ir/paper/78179/en

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