Reports of Biochemistry and Molecular Biology

en Construction of a Novel DNA Vaccine Candidate encoding LmSTI1-PpSP42 Fusion Protein from Leishmania major and Phlebotomus papatasi against Cutaneous Leishmaniasis زیست شناسی ملکولی Molecular Biology مقالات اصلی Original Article <div style="text-align: justify;">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. Conclusions: An LmSTI1Pp42 fusion protein was expressed HEK-293T cells. This construct may be an effective DNA vaccine against CL.  </div> Cloning, DNA vaccine, Leishmania major, LmSTI1, PpSP42. 67 75 http://rbmb.net/browse.php?a_code=A-10-139-1&slc_lang=en&sid=1 Touraj Miandoabi miandoabi.t@umsu.ac.ir 10031947532846006383 10031947532846006383 No Department of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Fariborz Bahrami f_bahrami@pasteur.ac.ir 10031947532846006384 10031947532846006384 No Pasteur Institute of Iran, Department of Immunology, 69 Pasteur Ave., Tehran, Iran. Vahideh Moein Vaziri vmvaziri@gmail.com 10031947532846006385 10031947532846006385 No Department of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Soheila Ajdary sohary@yahoo.com 10031947532846006386 10031947532846006386 Yes Pasteur Institute of Iran, Department of Immunology, 69 Pasteur Ave., Tehran, Iran.