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Title: Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus
Authors: Chaturvedi, Uttara
Kalim, Shahina
Desai, G
Ratta, Barkha
Kumar, Rajiv
Ravindra, P V
Kumar, Sudesh
Dash, B B
Tiwari, Sangeeta
Sahoo, A P
Tiwari, Ashok K
Keywords: Bicistronic gene construct
DNA vaccine
In vitro expression
Issue Date: Feb-2011
Publisher: NISCAIR-CSIR, India
Abstract: Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND
Description: 140-145
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.49(02) [February 2011]

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