Cloning and expression of recombinant flagellar protein flaB from Borrelia burgdorferi

OBJECTIVE: Lyme borreliosis is an arthropod transmitted infection caused by some species of the Borrelia genus. Current diagnosis employs serological testing and detection of Borrelia-specific antibodies. Using recombinant Borrelia burgdorferi antigens may improve assay specificity and sensitivity. One of the immunodominant Borrelia antigens that elicit a strong and early immune response is FlaB, making it appropriate for recombinant protein based serological diagnostic tests. MATERIAL AND METHODS: Borrelia burgdorferi genomic DNA was isolated and used as a template for the amplification of the flaB gene. The gene was cloned in the expression vector pGEX-2T. RESULTS: The amplified flaB gene was cloned in the expression vector yielding a GST-FlaB fusion gene. The gene ligated in-frame was expressed as the recombinant GST-FlaB protein. After visualization by polyacrylamide gel electrophoresis the successful expression of the FlaB protein was confirmed by immunoblotting. CONCLUSION: The expression and purification of the recombinant FlaB protein is a prerequisite for obtaining large amounts of the product through a simple and labour-free procedure, which will facilitate the diagnosis of Lyme disease.