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

Abstract

Background and Objectives: HBHA and Mtb32C have been isolated from culture supernatants of Mycobacterium tu- berculosis (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 (Mt- b32C-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 thiscell line. Conclusion: The constructed vector can produce two highly immunogenic antigens that fusion of them to gather makes chi- meric 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.

Keywords

• Mycobacterium tuberculosis
• HBHA
• Mtb32C
• Huh7.5 cells