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Expression of Leishmania major LmSTI1 in Yeast Pichia Pastoris

Mehdi Shokri, Soheila Ajdary, Mina Ebrahimi-Rad, Arash Memarnejadian, Mohamad-Hossein Alimohamadian, Fatemeh Motevali, Farzin Roohvand
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Abstract

Background: Leishmania major LmSTI1 is a conserved protein among different species of leishmania, and expressed in both amastigote and promastigote forms of L. major life cycle. It has previously been expressed in bacterial systems.

Materials and Methods: To express LmSTI1 in the methylotrophic yeast         Pichia pastoris (P. pastoris), the shuttle vector pPICZA containing gene lmsti1 was constructed under the control of the AOX1 promoter. The recombinant vector was electro-transformed into P. pastoris, and induced by 0.5% methanol in the buffered medium. The expression of the LmSTI1 protein was visualized in the total soluble protein of P. pastoris by 12% SDS-PAGE, and further confirmed by Western blotting with L.major-infected mouse sera and HRP-conjugated goat anti-mouse IgG as the first and secondary antibodies, respectively.

Results: The expression level was 0.2% of total soluble proteins.

Conclusion: It might be possible to use this formulation as a whole yeast candidate vaccine against cutaneous leishmanization.

Keywords

Pichia pastoris, LmSTI1, Leishmania major

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DOI: https://doi.org/10.22037/nbm.v5i1.11552