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Expression and characterization of immunodominant region of fusion protein of peste des petits ruminants virus in E. coli

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Title Expression and characterization of immunodominant region of fusion protein of peste des petits ruminants virus in E. coli
Not Available
 
Creator Apsana R
Balamurugan V
Veeragowda BM
Abraham S
Raju DSN
Rathnamma D
Byregowda SM
Rahman H
Shaila MS
 
Subject PPRV
Immunodominant ectodomain
F protein
E. coli
Expression
Purification
Immunoreactivity
 
Description Not Available
The present study envisages expression of immunodominant ectodomain of peste des petits ruminants
virus (PPRV) fusion (F) protein in Escherichia coli BL21 (DE3) and its characterization to assess
its immunoreactivity. The ectodomain gene sequences corresponding to 222 amino acids, was amplified
from PPR vaccine virus, cloned into pET33b vector and expressed in E. coli at an optimal temperature of
37 ◦C with 1 mM IPTG for 5 h. The expressed and Ni-NTA purified PPRV F protein (31 kDa) was characterized
by SDS-PAGE and Western blot using anti-his-tagged-conjugate, anti-serum raised against
recombinant PPRV F protein, hyper immune serum against whole PPRV and convalescent sera from sheep
and goats. The expressed protein was assessed for its immunoreactivity by ELISA and immunoblotting.
The antibody response mounted against the recombinant PPRV F protein in immunized rabbits was
detected by recombinant PPRV F antigen based indirect ELISA, and whole virus antigen based indirect
ELISA, which indicating the native confirmation ofthe expressed protein in E. coli. Indirect ELISA was optimized
using known true positive and negative sera with respect to PPRV antibodies in order to assess the
reactivity of the PPRV F protein in detecting PPRV F antibodies in small ruminants. The E. coli expressed
recombinant ectodomain of PPRV F protein exhibits immunoreactivity and was able to specifically detect
PPRV antibodies in response to both vaccination and disease in natural host.
Not Available
 
Date 2018-11-05T09:14:17Z
2018-11-05T09:14:17Z
2016-11-01
 
Type Research Paper
 
Identifier Not Available
0921-4488
http://krishi.icar.gov.in/jspui/handle/123456789/9336
 
Language English
 
Relation Not Available;
 
Publisher Elsevier