title: Cloning and Expression of Recombinant Plasmid Containing P36/LACK Gene of Leishmania infantum Iranian Strain.
abstract: BACKGROUND
  There are several methods, such as vaccination, to control visceral leishmaniasis. Although there is no efficient vaccine, it seem DNA vaccination with stimulates both cellular and humoral immunity apparently is the best way. The aim of this study was cloning and expression of LACK gene, a 36kD protein, as a candidate protein for vaccination against Iranian L. infantum.
  
  
  METHODS
  Iranian strain of L. infantum [MCAN/IR/07/Moheb-gh] was used as a template for PCR to amplify LACK gene. The LACK gene was cloned in pTZ57R/T vector and after confirmation it was digested by restriction enzymes (BamH1) and cloned in pcDNA3.1 expression vector. Recombinant plasmid was extracted and analyzed by sequencing, restriction digestion analysis and PCR reaction. The pc- LACK recombinant plasmid was purified from transformed E.coli (DH5α) and its expression was analyzed by SDS-PAGE and Western blot.
  
  
  RESULTS
  The results of sequencing, restriction digestion analysis and PCR reaction revealed that LACK gene was cloned correctly in pcDNA3.1 vector and the results of SDS PAGE and Western blot emphasized that LACK protein of Iranian L. infantum is a well-expressed protein.
  
  
  CONCLUSIONS
  We amplified, cloned and expressed Iranian L. infantum LACK gene successfully.
authors.affiliation: Dept. of Biology, Faculty of Basic science, Ahvaz branch, Islamic Azad University, Ahvaz, Iran ; Dept. of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
authors.forename: Saloomeh
authors.initials: S
authors.lastname: Shirali
authors.affiliation: Dept. of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
authors.forename: Hamidreza
authors.initials: H
authors.lastname: Haddadzadeh
authors.affiliation: Dept. of Medical Parasitology and Mycology, School of Public health, Tehran University of Medical Sciences, Iran ; Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohebali
authors.affiliation: Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Dept. of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
authors.affiliation: Dept. of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
authors.forename: Narges
authors.initials: N
authors.lastname: Amini
keywords: Cloning; Expression; LACK; Leishmania infantum; Visceral leishmaniasis