Record Details

Re-engineering of bicistronic plasmid pGPD/IFN to construct fusion gene co-expressing Glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) of <em>Edwardsiella tarda</em> and Interferon-gamma (IFN-γ) gene of <em>Labeo rohita</em> (Hamilton) and its <em>in vitro</em> functional analysis

Online Publishing @ NISCAIR

View Archive Info
 
 
Field Value
 
Authentication Code dc
 
Title Statement Re-engineering of bicistronic plasmid pGPD/IFN to construct fusion gene co-expressing Glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) of <em>Edwardsiella tarda</em> and Interferon-gamma (IFN-γ) gene of <em>Labeo rohita</em> (Hamilton) and its <em>in vitro</em> functional analysis
 
Added Entry - Uncontrolled Name Valsalam, Anisha
P, Gireesh-Babu
Rajendran, Kooloth Valappil
Tripathi, Gayatri
Bedekar, Megha Kadam
The funding of ICAR- Central Institute of Fisheries Education supported this work.
 
Uncontrolled Index Term DNA vaccine; Immune adjuvant; pcGPD/IFN; pIRES; Rohu
 
Summary, etc. Edwardsiella septicemia disease in the cultured Indian major carps is caused by the fish pathogen Edwardsiella tarda and it is preventable by DNA vaccination. Here, we tried to develop a bicistronic DNA vaccine pGPD/IFN expressing the Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene of Edwardsiella tarda and Interferon-gamma (IFN-γ) gene of Labeo rohita. The vaccine showed high protective efficiency in our previous studies; however as a limitation of bicistronic construct the expression of gene cloned in second frame (B) is poor. To overcome this limitation we re-engineered the construct and designed a fusion gene co-expressing the GAPDH and IFN-γ genes as one frame with an aim to get the optimum expression of both the genes. For this purpose, a fusion insert comprising GAPDH and IFN-γ coding sequences was cloned in to pcDNA3.1(+) plasmid vector. The fusion genes' in vitro expression was confirmed in the striped snakehead fish cell line (SSN-1). Successful expression of the re-engineered fusion gene DNA vaccine in the cell line was achieved at 48h post-transfection, which was confirmed by amplifying the expression transcripts of GAPDH and IFN-γ genes. Thus, the study concludes that the re-engineered fusion vaccine pcGPD/IFN (pcDNA3.1(+) plasmid having fusion GPD/IFN) is functional and can be effectively utilized to vaccinate rohu (Labeo rohita) as it contains the species-specific immune gene (IFN-γ) as an adjuvant.
 
Publication, Distribution, Etc. Indian Journal of Experimental Biology (IJEB)
2023-02-24 12:56:58
 
Electronic Location and Access application/pdf
http://op.niscair.res.in/index.php/IJEB/article/view/62517
 
Data Source Entry Indian Journal of Experimental Biology (IJEB); ##issue.vol## 61, ##issue.no## 03 (2023): IJEB [March 2023]
 
Language Note en