OVERPRODUCTION AND PURIFICATION OF MYCOBACTERIUM-TUBERCULOSIS CHAPERONIN-10

The overexpression of the Mycobacterium tuberculosis chaperonin 10 (Cp n10)-encoding gene was accomplished using baculovirus expression vecto rs. The product was immunoreactive with a Cpn10 monoclonal antibody (m Ab) and had an electrophoretic mobility identical to authentic Cpn10. The baculovirus system was most successful in terms of reaching nearly the full expression potential of the system. Recombinant Cpn10 was pu rified from recombinant baculovirus-infected Spodoptera frugiperda cel ls by isoelectrofocussing and size-exclusion chromatography. The bacul ovirus vector and purification methodology described represent a very powerful system for the large-scale production of the M. tuberculosis Cpn10 which may allow us to undertake structure-function analysis.