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Creation of leaderless FMDV replicon for development of replication defective virus (leaderless FMDV): A strategy towards the development of attenuated vaccine with marker facility

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Title Creation of leaderless FMDV replicon for development of replication defective virus (leaderless FMDV): A strategy towards the development of attenuated vaccine with marker facility
 
Creator Sarkar, Swaroop
Suryanarayana, V V S
Reddy, G R
Dechamma, H J
Shankar, S R Madhan
 
Subject Cloning
expression
Lb-protease deletion
T7 polymerase
FMDV virulence
BHK cells
attenuation
 
Description 26-34
Leader protease (Lpro) of foot-and-mouth disease virus (FMDV) which is essential for viral replication and pathogenicity
in host is found in two forms, Lab and Lb with similar activity but, both differing only at amino-termini with separate
initiation codons, 84 nucleotides apart. After translation of the genomic RNA into single polyprotein, the
L protease is released autocatalytic from the N terminus and cleaves the p220 subunit of the eukaryotic initiation factor 4F
complex resulting in the shut off host protein synthesis, an essential step for viral pathogenicity. We exploited this function
for development of attenuated virus by deleting the gene encoding Lb protease from FMDV Asia 1(63/72) cDNA replicon
(Joshi et al, 2013) by PCR mutagenesis approach. The deletions and intactness of the frames were confirmed by sequence
analysis. P1-2A polyprotein gene of FMDV ‘O’ was inserted into the replicon and the full length construct was studied for
virulence to baby hamster kidney (BHK) 21 cells. Vaccinia expressing T7 polymerase was used for in vitro RNA generation
and infection. The lower cytopathic effect (cpe) as observed by reduced replication efficiency confirmed the effect of Lb
deletion when compared with the construct with no deletion.
 
Date 2021-11-17T11:29:58Z
2021-11-17T11:29:58Z
2021-01
 
Type Article
 
Identifier 0975-0967 (Online); 0972-5849 (Print)
http://nopr.niscair.res.in/handle/123456789/58540
 
Language en
 
Publisher NIScPR-CSIR, India
 
Source IJBT Vol.20(1) [January 2021]