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Construction of a Novel DNA Vaccine Candidate encoding LmSTI1-PpSP42 Fusion Protein from Leishmania major and Phlebotomus papatasi against Cutaneous Leishmaniasis.


ABSTRACT: Background:Cutaneous leishmaniasis (CL) is a serious public health problem in many tropical countries. The infection is caused by a protozoan parasite of Leishmania genus transmitted by Phlebotominae sandflies. In the present study, we constructed a eukaryotic expression vector to produce a fusion protein containing LmSTI1 from Leishmania major (L. major) and PpSP42 from Phlebotomus papatasi (Ph. papatasi). In future studies we will test this construct as a DNA vaccine against zoonotic CL. Methods:The nucleotide sequences encoding the LmSTI1 protein and a fragment encoding 79% of PpSP42 were amplified using L. major and Ph. papatasi genomic DNA, respectively. The amplicons were cloned into the pcDNA3.1(+) eukaryotic expression vector. The recombinant plasmid pcDNA-LmSTI1Pp42 was propagated in Escherichia coli (E. coli) and used to transfect HEK-293T cells. The expressed fusion protein was analyzed by Western blotting using anti-LmSTI1 mouse serum. Results:Sequences encoding LmSTI1 and partial PpSP42 were cloned into pcDNA3.1(+). Production of the recombinant LmSTI1Pp42 fusion protein was confirmed by Western blotting. Conclusion:An LmSTI1Pp42 fusion protein was expressed HEK-293T cells. This construct may be an effective DNA vaccine against CL.

SUBMITTER: Miandoabi T 

PROVIDER: S-EPMC6175588 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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Construction of a Novel DNA Vaccine Candidate encoding LmSTI1-PpSP42 Fusion Protein from <i>Leishmania major</i> and <i>Phlebotomus papatasi</i> against Cutaneous Leishmaniasis.

Miandoabi Touraj T   Bahrami Fariborz F   Moein Vaziri Vahideh V   Ajdary Soheila S  

Reports of biochemistry & molecular biology 20181001 1


<h4>Background</h4>Cutaneous leishmaniasis (CL) is a serious public health problem in many tropical countries. The infection is caused by a protozoan parasite of Leishmania genus transmitted by Phlebotominae sandflies. In the present study, we constructed a eukaryotic expression vector to produce a fusion protein containing LmSTI1 from <i>Leishmania major (L. major)</i> and PpSP42 from <i>Phlebotomus papatasi (Ph. papatasi)</i>. In future studies we will test this construct as a DNA vaccine agai  ...[more]

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