title: Expression and Purification of Recombinant ROP1 of Toxoplasma gondii in Bacteria.
abstract: BACKGROUND
  Toxoplasmosis is a worldwide-distributed infection which is mostly asymptomatic but can cause serious health problems in congenitally-infected newborns and immunecompromised individuals. Research is undergoing both to improve Toxoplasma serological tests, which play the main role in laboratory diagnosis of the infection, and develop an effective vaccine to prevent the infection. Some studies showed usefulness of rhoptry protein 1 (ROP1) antigen of Toxoplasma gondii (T. gondii) in serodiagnosis of the infection and induction of protective immunity. The purpose of this study was to produce recombinant ROP1 and evaluate its antigenicity against human infected sera.
  
  
  METHODS
  DNA encoding ROP1, amino acids 171 to 574, was obtained from T. gondii RH strain by polymerase chain reaction amplification and cloned in prokaryotic expression plasmid pET-15b. rROP1 was expressed in Escherichia coli (E. coli) and purified in a single step by immobilized metal ion affinity chromatography.
  
  
  RESULTS
  DNA sequencing showed 99% similarity between the cloned sequence and the corresponding sequence in Gene bank. Results indicated the proper antigenicity of rROP1. Sera from Toxoplasma infected individuals specifically recognized rROP1 in Western blotting.
  
  
  CONCLUSIONS
  rROP1 is antigenic toward human infected sera and can be used in studies for development of both a Toxoplasma serological test and a protective vaccine.
authors.affiliation: Molecular Parasitology Laboratory, Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran ; Department of Biochemistry, Science and Research Branch, Islamic Azad University, Fars, Iran.
authors.forename: Reyhaneh
authors.initials: R
authors.lastname: Mohabati
authors.forename: Jalal
authors.initials: J
authors.lastname: Babaie
authors.forename: Samira
authors.initials: S
authors.lastname: Amiri
authors.forename: Mohammad
authors.initials: M
authors.lastname: Talebzadeh
authors.forename: Pezhman
authors.initials: P
authors.lastname: Fard-Esfahani
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Darbouy
authors.forename: Majid
authors.initials: M
authors.lastname: Golkar
keywords: Gene expression; Purification; ROP1 protein; Toxoplasma gondii