화학공학소재연구정보센터
Biochemical and Biophysical Research Communications, Vol.282, No.5, 1183-1188, 2001
Cloning and expression of human rotavirus spike protein, VP8*, in Escherichia coli
A system for the expression and purification of soluble VP8*, part of the human rotavirus (HRV) spike protein, was established by expressing VP8* as a fusion protein with glutathione S-transferase (GST). VP8 cDNA, from the Wa strain of HRV, was prepared by RT-PCR, cloned into a pUC18 plasmid, and inserted into a pGEXi-4T-2 GST fusion vector. The GST-VP8* fusion protein was expressed in Escherichia coli, and the VP8* was purified by Glutathione Sepharose 4B affinity chromatography, yielding 1.8 mg VP8*/L culture. The purified VP8* was used to vaccinate chickens, eliciting antibodies which display-ed high neutralization activity against the Wa strain of HRV, suggesting its use for the induction of specific neutralizing antibodies for potential immunotherapentic applications for the prevention of HRV infection.