[Expression and purification of dormancy survival regulator Rv2628 of Mycobacterium tuberculosis and preparation of rabbit anti-Rv2628 polyclonal antibody].

Objective To express and purify the dormancy survival regulator Rv2628 of Mycobacterium tuberculosis, so as to prepare and identify its rabbit polyclonal antibody. Methods Using the genome of Mycobacterium tuberculosis H37Rv strain as a template, the Rv2628 gene was amplified to construct a recombinant expression plasmid which was transformed into an Escherichia coli protein expression strain and induced expression by IPTG. The target protein was purified using Ni-NTA chromatography column, and the rabbit polyclonal antibody was obtained by immunizing New Zealand white rabbits with purified Rv2628 protein. The specificity of polyclonal antibodies was verified by Western blot analysis and indirect ELISA, respectively. Results The PET-30A-RV2628 recombinant carrier was successfully constructed. After induction by IPTG, the RV2628 protein was mainly expressed in the form of inclusion. The high-purity Rv2628 protein was obtained by Ni-NTA column purification. Rabbit anti-Rv2628 polyclonal antibody was obtained after immunizing the rabbit. The antibody had good antigen binding properties and the antibody titer reached 1:1 093 500. Conclusion The high-purity Rv2628 protein and high-titer rabbit anti-Rv2628 polyclonal antibody were successfully prepared.