Construction Of Broad Host Range Vector For Expression Of Heterologus Protein In Brucella Abortus Strain 19 | 4729
Journal of Biotechnology & Biomaterials
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Brucellosis, a major zoonotic disease causing abortion and infertility in cattle, is controlled mainly by vaccination with strain
Brucella abortus strain 19. It is a smooth, attenuated and live vaccine candidate which can induce a strong humoral and
cellular immune response and therefore, it is an attractive vector platform for the delivery of heterologus antigens. The objective
of this study was to construct a broad host range vector for expression of heterologus protein in B abortus strain 19. For this,
promoter sequence of Brucella groE gene was cloned into broad host range vector pBBR 122. For one step purification and easy
detection of the expressed protein, six histidine tag was added in to the reverse primer of groE promoter. To check promoter
activity and expression status of the construct, a promoterless GFP gene was cloned into the vector. The construct pBBR GroE_
GFP was then electroporated in to strain 19. The expression of GFP was induced by heat shock at 42
C and treatment with H
Then the recombinant protein was purified by using urea lysis method and followed by Ni-NTA affinity chromatography. Further,
the purified protein was dialysed and the specific reactivity of the recombinant protein was checked by western blotting using
anti histidine antibody. A single specific band could be detected at 28kDa confirming the expressed recombinant protein. Based
on the present study, it is concluded that B abortus strain 19 can be used as vector for expression of foreign protein and further
experimentation is required study its vaccinal potential.
Justin Davis K is the native of Kerala, India. He has completed his MVSc degree from Indian Veterinary research institute in veterinary epidemiology
and has procured PhD from the same institute on Veterinary Bacteriology. He had received IVRI fellowship for both MVSc and PhD. He had also
received PTF grant for young scientists for presenting a paper at Estonia conference.
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