Brucellosis is an important and re-emerging zoonotic disease worldwide. The prevention of human infection is achieved predominantly through the control of brucellosis in agricultural animals, which in turn depends on accurate diagnosis and vaccination. However, conventional serological diagnosis of brucellosis has several limitations, and currently available vaccines for animals have several drawbacks, including the ability to cause infection in humans. Phosphoglycerate kinase (Pgk) is one of the specific proteins reactive with mouse sera in the early stage of Brucella infection, and deletion of the pgk gene in B. abortus strain 2308 resulted in extreme attenuation of this strain in vitro and in vivo. Furthermore, the B. abortus pgk mutant has been used as a live vaccine, and in challenge experiments, it induced protection that was superior to that conferred by commercial strains. In this study, the pgk gene from Brucella abortus 544 was successfully amplified and cloned into a maltose binding protein fusion protein expression vector (pMAL). The recombinant protein was expressed in Escherichia coli DH5α and purified. The immunogenicity of purified recombinant B. abortus 544 Pgk (rPgk) was evaluated by western blot analysis using Brucella-positive mouse sera. rPgk could be used as an antigenic component for future serological tests and potential vaccine development.

Abstract
 Introduction
 Materials and Methods
  Bacterial strains and growth conditions
  Primer design and PCR amplification
  Cloning, expression, induction, and purification of rPgk in the pMAL vector
  Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting
 Results and Discussion
 References

• Alisha Wehdnesday Bernardo Reyes(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, College)
• Hannah Leah Tadeja Simborio(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
• HuynhTanHop(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
• Lauren Togonon Arayan(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
• Wongi Min(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
• Hu Jang Lee(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
• Hong Hee Chang(Institute of Agriculture and Life Science, Gyeongsang National University)
• Suk Kim(Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Institute of Agriculture and Life Science, Gyeongsang National University) Corresponding Author