cloning and expression of soluble recombinant hiv-1 crf35 protease-hp thioredoxin fusion protein

background: as a drug target and an antigenic agent, hiv-1 protease (hiv-1 pr) is at the center of attention for designing anti-aids inhibitors and diagnostic tests. in previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in e. coli and investigation of its immunoreactivity. methods: protease coding region was isolated from the serum of an infected individual, amplified by rt-pcr and cloned into ptz57r using ta-cloning. protease coding frame was isolated by pcr and cloned in pet102/d. topo expression vector and cloned protease was expressed in escherichia coli (e. coli) bl21. produced recombinant protein was purified by affinity ni-nta column and protein concentration was checked by bca protein assay kit. subsequently, immunoreactivity of recombinant protease (rpr) was assayed by western blotting and elisa. results: cloning of the hiv protease by topo cloning system in pet102/d.topo was confirmed with pcr and sequencing. the concentration range of purified recombinant protein was 85 to 100 μg/ml. immunogenicity of rpr was confirmed by western blotting and elisa. conclusion: soluble production of recombinant hiv-1 protease (hiv-1 rpr) was performed successfully. this recombinant protein disclosed 86% specificity and 90% sensitivity in immunoassay tests.