Comparison on different extraction protocols of PK-15 cell membrane proteins and identification of cellular receptors for classical swine fever virus

The aim of this study was to compare the effects of four methods,Mem-PER Eukaryotic Membrane Protein Extraction Reagent Kit,freezing-thaw,ultrasonic treatment and low permeability cracking method,on extracting PK-15 cell membrane proteins.The results showed that ultrasonic treatment was more ideal for the extraction of membrane proteins in PK-15 cell.After SDS-polyacrylamide gel electrophoresis(SDS-PAGE),the PK-15 cell membrane proteins were transferred to nitrocellulose membrane,and virus overlay protein binding assay(VOPBA) was performed to identify the receptor protein for classical swine fever virus(CSFV).There were three cellular proteins of 90 ku to 100 ku that could bind CSFV in PK-15 cells,indicating that these proteins might be putative receptors or a part of receptor complex for CSFV.