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Title: Expression of Capsid Precursor Polypeptide (PI) of Foot and Mouth Disease Virus Vaccine Strains in Pichia pastoris
Authors: Renji, R
Balamurugan, V
Saha, S N
Reddy, G R
Suryanarayana, V V S
Keywords: FMDV;Serotypes;P1polyprotein;Pichia pastoris;Expression
Issue Date: Oct-2003
Publisher: NISCAIR-CSIR, India
Abstract: Foot and mouth disease virus (FMDV) is the aetiological agent of a highly contagious vesicular disease of c1ovenhooved animals. The gene coding for the capsid polyprotein (P1) of FMDV from vaccine strains was amplified and cloned into the Yeast transfer vector (pPIC-9K) at Eco RI and Not I. The cloned PI gene was characterized by colony PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid of each serotype was transferred into GS1l5 strain of Pichia by electroporation. The His+ Pichia transformants were anaIysed for the presence of the insert in the yeast genome by PCR. PCR positive clones of each serotype were grown separately and expression was induced with 0.5% methanol. The expressed gene product in yeast, Pichia pastoris was characterized by SDS-PAGE and Western Blot analysis. This is the first report on the production of FMDV structural proteins in yeast.
Page(s): 533-538
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.02(4) [October 2003]

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