Evaluation of the eukaryotic expression of mtb32C-hbha fusion gene of Mycobacterium tuberculosis in Hepatocarcinoma cell line.

BACKGROUND AND OBJECTIVES: HBHA and Mtb32C have been isolated from culture supernatants of Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and their immunogenicity previously studies have been confirmed. In this study, capability of constructed vector containing two mycobacterial immunodaminant antigens (Mtb32C-HBHA), in producing new chimeric protein under the in-vitro condition was examined.

MATERIALS AND METHODS: In present study Huh7.5 cells was transfected with Mtb32C-HBHA -pCDNA3.1+ recombinant vector using the calcium phosphate method and expression of chimeric protein was assessed by RT-PCR and Western blot methods.

RESULTS: Results of RT-PCR and Western blot showed expression of 35.5 KD recombinant protein (Mtb32C-HBHA) in this cell line.

CONCLUSION: The constructed vector can produce two highly immunogenic antigens that fusion of them to gather makes chimeric antigen with new traits. Other attempts are needed to evaluate specific properties of this new antigen such as molecular conformation modeling and immunologic characteristics in future studies.