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Expression of Brucella Iumazine synthase gene in E. coli system.

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Title Expression of Brucella Iumazine synthase gene in E. coli system.
Not Available
 
Creator Nagalingam M
Thaslim J B
Vijaya Gowri N
Balamurugan V
Shome R
Manjunatha Reddy G B
Shome B R
Rahman H
Roy P
Gandham R K
 
Subject Brucellosis
Brucella Lumazine Synthase (BLS)
Recombinant protein
TOPO vector
 
Description Not Available
Brucellosis is an important zoonosis of serious economic and public health consequences
affecting livestock and human beings caused by Brucella spp. The available whole cell
antigen and smooth lipopolysaccharide (sLPS) based diagnostics show false positive
reactions due to cross reactivity. Therefore, in this study, Brucella lumazine synthase
(BLS), an immunogenic protein of Brucella spp. coding 477 bp bls gene was amplified
from DNA extracted from Brucella abortus S99 strain and the amplified product was
cloned in pET directional TOPO expression vector and transformed in E. coli TOP 10F’
cells. After verifying sequence, bls recombinant clones were subsequently transformed in
E. coli BL21 (DE3)/pLysS cells for expression. BLS protein expression was induced with
1mM IPTG and optimized, six hours induction yielded maximum expression of BLS
protein. The expressed protein was purified by His-tag affinity purification method using
Ni-NTA and characterized by SDS-PAGE and Western blot. Recombinant BLS protein
expressed in E. coli has potential use as diagnostic antigen or immunogen for development
of diagnostics and vaccine for brucellosis
Not Available
 
Date 2018-11-12T05:07:53Z
2018-11-12T05:07:53Z
2017-10-10
 
Type Research Paper
 
Identifier Not Available
2319-7706
http://krishi.icar.gov.in/jspui/handle/123456789/10258
 
Language English
 
Relation Not Available;
 
Publisher Excellent publishers