title: High-Level Expression of Immunogenic Recombinant Plasmodium vivax Merozoite Surface Protein (Pvmsp-142 kDa) in pGEX 6P1 Vector.
abstract: BACKGROUND
  Detection of Plasmodium vivax specific antibodies with serological tests could be a valuable tool for epidemiological researches. Whereas P. vivax cannot be simply obtained in vitro, serological tests using total or semi-purified antigens are infrequently used. Given this restriction, the present study investigated whether recombinant P. vivax merozoite surface protein 1 (PvMSP-1 42 kDa) could be useful in detection of antibodies from the serums of a P. vivax infected person using serological tests.
  
  
  METHODS
  Parasite DNA was extracted from blood sample of an Iranian P. vivax-infected patient. The region of PvMSP-142 kDa was amplified by PCR then cloned into pTZ57R/T vector and sequenced. The insert was sub cloned into pGEX 6P1 expression vector. Afterwards, it was transformed into E. coli BL21 and cultured massively. Sub cloning of gene was confirmed by PCR and enzyme digestion and sequencing finally. Production of recombinant protein was confirmed by SDS-PAGE. Western blot was performed by human sera to appraisal binding ability to the IgG antibodies of P. vivax infected patients. Recombinant protein was purified and estimated by Bradford assay.
  
  
  RESULTS
  The specialty values of the Western blot determined with 10 sera from naturally infected individuals, 10 sera from healthy individuals and 7 sera from individuals with other infectious diseases.
  
  
  CONCLUSIONS
  For the Iranian population, using a Western blot assay for MSP-142 recombinant protein can be used as the foundation for promotion of serological assay for the detection of P. vivax malaria such as ELISA.
authors.affiliation: 1. Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences , Tehran, Iran . ; 2. Dept. of Parasitology and Mycology, Zahedan University of Medical Sciences , Zahedan, Iran.
authors.forename: Hadi
authors.initials: H
authors.lastname: Mirahmadi
authors.affiliation: 3. Dept. of Parasitology and Mycology, Lorestan University of Medical Sciences , Khoramabad, Iran.
authors.forename: Shirzad
authors.initials: S
authors.lastname: Fallahi
authors.affiliation: 1. Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences , Tehran, Iran .
authors.forename: Vahid
authors.initials: V
authors.lastname: Fallah Omrani
authors.affiliation: 1. Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences , Tehran, Iran .
authors.forename: Bahram
authors.initials: B
authors.lastname: Kazemi
authors.affiliation: 1. Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences , Tehran, Iran .
authors.forename: Ali
authors.initials: A
authors.lastname: Haghighi
authors.affiliation: 1. Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences , Tehran, Iran .
authors.forename: Seyyed Javad
authors.initials: SJ
authors.lastname: Seyyed Tabaei
keywords: Expression vector; Iran; Plasmodium vivax; Recombinant PvMSP-142 kDa