Reports of Biochemistry and Molecular Biology
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Basic Sciences
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2322-3480
2322-3480
10.61186/rbmb
en
jalali
1397
7
1
gregorian
2018
10
1
7
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en
Construction of a Novel DNA Vaccine Candidate encoding LmSTI1-PpSP42 Fusion Protein from <em>Leishmania major</em> and <em>Phlebotomus papatasi</em> against Cutaneous Leishmaniasis
زیست شناسی ملکولی
Molecular Biology
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background: </em></strong>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<em> Leishmania major (L. major)</em> and PpSP42 from <em>Phlebotomus papatasi (Ph. papatasi)</em>. In future studies we will test this construct as a DNA vaccine against zoonotic CL.<br>
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<strong><em>Methods:</em></strong> The nucleotide sequences encoding the LmSTI1 protein and a fragment encoding 79% of PpSP42 were amplified using <em>L. major </em>and <em>Ph. papatasi</em> genomic DNA, respectively. The amplicons were cloned into the pcDNA3.1(+) eukaryotic expression vector. The recombinant plasmid pcDNA-LmSTI1Pp42 was propagated in <em>Escherichia coli (E. coli)</em> and used to transfect HEK-293T cells. The expressed fusion protein was analyzed by Western blotting using anti-LmSTI1 mouse serum.<br>
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<strong><em>Results:</em></strong> Sequences encoding LmSTI1 and partial PpSP42 were cloned into pcDNA3.1(+). Production of the recombinant LmSTI1Pp42 fusion protein was confirmed by Western blotting.<br>
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<strong><em>Conclusions:</em></strong> An LmSTI1Pp42 fusion protein was expressed HEK-293T cells. This construct may be an effective DNA vaccine against CL.<br>
</div>
Cloning, DNA vaccine, Leishmania major, LmSTI1, PpSP42.
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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.