Title: Expression study of recombinant p1 protein of local foot and mouth disease virus in yeast cells

Abstract:

Foot and mouth disease is one of the most lethal viral animal diseases worldwide in cleft hoofed animals such as cattle’s, sheep, water buffalo etc. The virus belongs to family Picornaviridae and genus Apthovirus. There are seven serotypes (A, O, C Asia1, and SAT 1-3) of FMDV exist. The serotype O is the most dominant disease in Pakistan. The foot and mouth disease virus protein shell comprises the P1 polyprotein. The P1 polyprotein cleaves into four virus structural proteins VP4, VP3, VP2, and VP1. Among these the VP4 is entirely internal. The capsid protein P1 is associated with the antigenic properties and interact with cell surface receptors.

Inactivated virus vaccine for the prevention of FMD has been commercialized. Vaccination is an only way to overcome this virus. Pichia pastoris is a methylotrophic yeast and possess a strong inducible promoter AOX1.it is a very cost effective eukaryotic expression system with the benefit of post translational modification which is necessary for the proper confirmation of the protein. This study aims to clone the P1 polyprotein gene of local FMDV serotype O strain in methylotrophic yeast Pichia pastoris expression system.

For this purpose P1 gene was modified in Pichia pastoris and copy number was increased in E. coli. The synthetic gene was expressed in P. pastoris GS115 expression system to produce protein. Protein expression was analyzed by using SDS-PAGE and Western Blot. Protein was quantified by using Bradford assay. Indirect ELISA was done to check the expression of the P1 polyprotein. FMDV serotype ‘O’ builted 96 well plates after saturation were used for trapping the antibody. This study would be helpful in construction of a recombinant expression system containing P1 gene from local FMDV (serotype O) in a eukaryotic P. pastoris expression system which will be further used to study the expression of viral protein and to produce recombinant P1 polyprotein as a candidate antigen for the production of recombinant vaccine against FMDV  

Biography:

Muhammad  Asif  Muneer  from University of Veterinary and Animal Sciences Lahore, Pakistan. With full command on High pressure liquid chromatography, solid phase extraction assembly , spectrophotometer, thin layer chromatography and PCR.