title: Cloning and expression ofClostridium perfringenstype D vaccine strain epsilon toxin gene inE. colias a recombinant vaccine candidate.
abstract: OBJECTIVE
  Clostridium perfringens,a Gram-positive obligate anaerobic bacterium, is able to form resistant spores which are widely distributed in the environment.C. perfringensis subdivided into five types A to E based on its four major alpha, beta, epsilon and iota toxins. The aim of the present study was cloning and expression ofC. perfringenstype D vaccine strain epsilon toxin gene.
  
  
  METHODS
  Genomic DNA was extracted and the epsilon toxin gene was amplified usingPfuDNA polymerase. The PCR product was cloned into pJET1.2/blunt cloning vector. The recombinant vector (pJETε) was sequenced using universal primers. At the next step epsilon toxin gene was subcloned into pET22b(+) expression vector and transformed intoE. coliRosetta (DE3) host strain.
  
  
  RESULTS
  The recombinant protein has been expressed inE. coliRosetta (DE3) cells after subcloning ofC. perfringens etxgene (1008 bp) into the expression vector.
  
  
  CONCLUSIONS
  We concluded thatE. coliRosetta strain was suitable for the expression of recombinantC. perfringensepsilon toxin protein from pET22ε expression vector. This recombinant cell can be used for further research on recombinant vaccine development.
authors.affiliation: Islamic Azad University, Karaj Branch, Department of Microbiology, Karaj, Iran.
authors.forename: Parastoo
authors.initials: P
authors.lastname: Aziminia
authors.affiliation: Clostridia Specialized Research Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Pilehchian-Langroudi
authors.affiliation: Razi Reference Laboratory of Tileriosis Research, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
authors.forename: Kasra
authors.initials: K
authors.lastname: Esmaeilnia
keywords: C. perfringens; Cloning; Epsilon toxin gene; Expression; Vaccine strain