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

Simple item page
dc.contributor.authorValsalam, Anishaen_US
dc.contributor.authorP, Gireesh-Babuen_US
dc.contributor.authorRajendran, Kooloth Valappilen_US
dc.contributor.authorTripathi, Gayatrien_US
dc.contributor.authorBedekar, Megha Kadamen_US
dc.date.accessioned2023-08-09T06:21:17Z
dc.date.available2023-08-09T06:21:17Z
dc.date.issued2023-03
dc.description.abstractEdwardsiella 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-?) 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-? 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-? 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-? 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-?) as an adjuvanten_US
dc.identifier.affiliationsDepartment of Aquatic Animal Health Managementen_US
dc.identifier.affiliationsDepartment of Fish Genetics and Biotechnology, ICAR- Central Institute of Fisheries Education, Mumbai 400 061, Indiaen_US
dc.identifier.affiliationsPresent Address: ICAR- National Research Centre on Meat, Hyderabad-500 092, Indiaen_US
dc.identifier.citationValsalam Anisha, P Gireesh-Babu, Rajendran Kooloth Valappil, Tripathi Gayatri, Bedekar Megha Kadam. Re-engineering of bicistronic plasmid pGPD/IFN to construct fusion gene co-expressing Glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) of Edwardsiella tarda and Interferon-gamma (IFN-?) gene of Labeo rohita (Hamilton) and its in vitro functional analysis. Indian Journal of Experimental Biology. 2023 Mar; 61(3): 204-213en_US
dc.identifier.issn0975-1009
dc.identifier.issn0019-5189
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/222588
dc.languageenen_US
dc.publisherCSIR-NIScPRen_US
dc.relation.issuenumber3en_US
dc.relation.volume61en_US
dc.source.urihttps://doi.org/10.56042/ijeb.v61i03.62517en_US
dc.subjectDNA vaccineen_US
dc.subjectImmune adjuvanten_US
dc.subjectpcGPD/IFNen_US
dc.subjectpIRESen_US
dc.subjectRohuen_US
dc.titleRe-engineering of bicistronic plasmid pGPD/IFN to construct fusion gene co-expressing Glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) of Edwardsiella tarda and Interferon-gamma (IFN-?) gene of Labeo rohita (Hamilton) and its in vitro functional analysisen_US
dc.typeJournal Articleen_US
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
IJEB2023v61n3p204.pdf
Size:
1.16 MB
Format:
Adobe Portable Document Format
Download
Collections
Indian Journal of Experimental Biology