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Identification of Porcine haemagglutinating encephalomyelitis virus receptor in PK cell membranes

Published online by Cambridge University Press:  27 June 2008

Lu Hui-Jun ,
He Wen-Qi ,
Song De-Guang ,
Liu Li-Guo ,
Chang Ling-Zhu ,
Li Zhi-Ping ,
Chen Ke-Yan  and
Gao Feng
Lu Hui-Jun
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
He Wen-Qi
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Song De-Guang
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Liu Li-Guo
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Chang Ling-Zhu
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Li Zhi-Ping
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Chen Ke-Yan
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Gao Feng*
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

To identify Porcine haemagglutinating encephalomyelitis virus (HEV) 67N receptor in porcine kidney (PK) cell membranes, the S1 protein of HEV was expressed in Pichia pastoris and purified by Ni2+ affinity chromatograph. Polyclonal antibodies to HEV were prepared by immunizing rabbits by injecting the purified S1 protein four times. After SDS–polyacrylamide gel electrophoresis (SDS–PAGE), the PK cell membrane proteins were transferred on to nitrocellulose membrane. A virus overlay protein binding assay (VOPBA) was performed using the recombinant S1 protein to identify the protein binding receptor, HEV-S1. The result showed that HEV-S1 protein bound to one band (about 90 kDa) in PK cell membranes. This result is very important for the study of the pathogenic mechanism of HEV.

Keywords

Porcine haemagglutinating encephalomyelitis virusvirus overlay protein binding assayvirus receptor
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 1 , April 2008 , pp. 87 - 92
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(1): 20–23

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