Cloning and Recombinant Expression of Elongation Factor-1α of Leishmania mexicana

Vector Borne Zoonotic Dis. 2018 Dec;18(12):709-712. doi: 10.1089/vbz.2018.2347. Epub 2018 Sep 27.

Abstract

Leishmania mexicana is an intracellular parasite that causes cutaneous leishmaniasis (CL) in some countries, including Mexico. Recently, we identified the elongation factor-1α (EF-1α) of L. mexicana by immunoproteomic analysis. In Leishmania donovani, this molecule has been reported as a virulence factor involved in downregulation of macrophages by no-canonical function when EF-1α interacts with protein tyrosine phosphatase-1 (SHP-1). However, in L. mexicana the key role of EF-1α in host-parasite relationship has not been elucidated, by this reason we started with cloning and recombinant expression of this antigen. A sequence of 1350 bp corresponding to EF-1α (EF-Lm) full-length gene was amplified from genomic DNA of L. mexicana (GenBank: MG256973); this gene contains only one nucleotide change: C464T, compared with L. mexicana reference sequence (GenBank: FR799570.1). The gene cloned (EF-Lm) codes for a protein of 449 residues. It was expressed in Escherichia coli and purified as 63 kDa sumo-fusion protein, which was recognized in the sera of patients diagnosed with CL. Our results show that EF-Lm is immunogenic during infection, and the rEF-Lm could be used for further analyses in the host-parasite relationship.

Keywords: Leishmaniasis; elongation factor-1α; recombinant protein.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular*
  • DNA, Helminth
  • Gene Expression Regulation
  • Leishmania mexicana / genetics
  • Leishmania mexicana / metabolism*
  • Peptide Elongation Factor 1 / genetics
  • Peptide Elongation Factor 1 / metabolism*

Substances

  • DNA, Helminth
  • Peptide Elongation Factor 1