Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar-243122, (U.P.), India
Online published on 19 December, 2013.
Black quarter is an important disease of ruminants caused by Clostridium chauvoei, a Gram-positive, anaerobic, spore-forming bacterium causing serious toxemia and high mortality in livestock. NAD-dependent beta-hydroxybutyryl-CoA dehydrogenase (HBHD) is an immunogenic cell-surface associated protein of C. chauvoei and has the potential to be a vaccine candidate against C. chauvoei infections. The 3-hydroxybutyryl-CoA dehydrogenase was identified as an immunogenic protein molecule of Flavobacterium columnare and it may provide candidate molecules for the development of vaccines against columnaris disease in fish. In the present study, the NAD-dependent HBHD gene of C. chauvoei was amplified by PCR (844bp) and cloned into expresso rhamnose SUMO vector, transformed into E. cloni 10G chemically competent cells and expression was induced with L-rhamnose. Screening of the induced colonies by SDS-PAGE analysis revealed an expected protein band of about 43kDa size. The expression of recombinant protein was also confirmed by Western blotting. Expressed recombinant HBHD protein was purified and will be used for evaluation of its protective efficacy.
