co-expression of hbha and mtb32c genes from mycobacterium tuberculosis h37rv in a prokaryotic system

Background: heparin-binding hemagglutinin (hbha) protein is a surface adhesin that mediates the attachment of mycobacterium tuberculosis to host cells by its own unique, carboxyl-terminal region. the methylated hbha has a specific motif with a lysine-, alanine-, and proline-rich domain. more recently, it has been shown that hbha protein has potential activity in stimulating immune responses, and is a promising new candidate for diagnostic applications besides a protective antigen against tuberculosis. objectives: the aim of this study was to isolate a mycobacterial latency gene (hbha), and subsequently produce its protein as a new antigen for the interferon-gamma release assay test (igras). methods: in the present work, hbha and mtb32c genes were isolated from the mycobacterium tuberculosis h37rv genome using the polymerase chain reaction (pcr) method. the pcr products and pet21+ vector were digested with specific restriction enzymes and then submitted to the ligation procedure. escherichia coli bl21-codonplus (de3) competent cells were transformed with the recombinant mtb32c-hbha -pet21+ vector. expression of recombinant protein (mtb32c-hbha) was confirmed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page) and western blot methods. results: detection of a 500-bp gene and sequencing of recombinant pet-mtb32c-hbha vector, all confirmed the accuracy of the cloning procedure. a 36-kda band of mtb32c-hbha protein was also detected by western blotting. conclusions: in this study, expression of mtb32c-hbha protein was successfully done, in the prokaryotic system. further studies are needed to evaluate the efficacy of recombinant mtb32c-hbha protein in diagnosis of latent tuberculosis.