Production of recombinant HTLV1 Tax protein in Rosetta(DE3) bacterial host

HTLV-1 is the first declared Retrovirus that cause disease in human. physio pathology of HTLV-1 related diseases, was linked with tax protein characteristic.Use of mutant Tax proteins accompanied by immune regulator drugs could help treating HTLV1 Associated Myelopathy patients as a modulator of potent immune response against this viral protein.Since Tax protein is not commercially available, produce of recombinant Tax protein was targeted for this study. Matherial and Coding sequence of Tax protein (contain R222K mutant) in the pcDNA3.1(+) digested with BamHI and XhoI restriction enzymes then removed and then inserted into the expression vector pET32a(+) within the same cutting site s and cloned into Ecoli DH5α. Recombinant vector was confirmed with enzyme digestive processes,colony PCR and sequencing of cloned gene and then expression vector. E-coli Rosetta (DE3) was transformed with the recombinant plasmid and the expression was induced. The expression of protein was assayed with SDS-PAGE and Western blot. Using monoclonal antibodies against Tax and 5His epitope. Finally antigenic characteristic of the recombinant protein was evaluated by wester n blotting against patient serums. presence of Tax protein band in the recording of SDS-PAGE and Westernblot was confirmed.western blotting the recombinant protein with patient sera showed the band related to Tax protein. The expression recombinant protein is well produced and could be detected by patient's sera,making it eligible to be used as a recombinant viral antigen for future purposes.