title: Characterization and expression of e2 glycoprotein of classical Swine Fever virus in a eukaryotic expression system.
abstract: Classical swine fever (CSF) is an economically important Office International des Epizooties (OIE) list A disease of swine characterized by high fever and multiple haemmorhages. The E2 glycoprotein of CSFV is immunogenic and induces neutralizing antibodies against CSFV. In the present study, complete coding region of the E2 gene from Indian virulent field isolate (Mathura) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subsequently cloned into a mammalian expression vector; pcDNA3.1(+) at BamHI and XbaI site. The recombinant plasmid; pcDNA.E2.CSFV. was confirmed by restriction enzyme digestion. The pcDNA.E2.CSFV. transfected Vero cell expressed E2 protein which was confirmed by western blotting, immunoperoxidase and indirect immunofluorescent tests. Additionally, flow cytometry analysis also confirmed that 15% of transfected Vero cells expressed the E2 glycoprotein compared to mock or vector alone transfected cells. Further study is under way to evaluate recombinant pcDNA.E2.CSFV. Mathura clone as DNA vaccine against CSFV.
authors: Immuno-Chemistry Laboratory, Department of Biochemistry, Indian Veterinary Research Institute, Izatnagar, 243122 UP India.
authors: Barkha
authors: B
authors: Ratta
authors: Binita
authors: B
authors: Nautiyal
authors: P V
authors: PV
authors: Ravindra
authors: Uttara
authors: U
authors: Chaturvedi
authors: Sudesh
authors: S
authors: Kumar
authors: P K
authors: PK
authors: Subudhi
authors: Kantaraja
authors: K
authors: Chindera
authors: Sangeeta
authors: S
authors: Tiwari
authors: N N
authors: NN
authors: Barman
authors: Ashok K
authors: AK
authors: Tiwari
keywords: Classical swine fever (CSF); Classical swine fever virus (CSFV); Reverse transcription-polymerase chain reaction (RT-PCR)
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