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 h uman 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 in serted 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 Se pharose 4B affinity chromatography, yielding 1.8 mg VP8*/L culture. The pur ified VP8* was used to vaccinate chickens, eliciting antibodies which displ ay-ed high neutralization activity against the Wa strain of HRV, suggesting its use for the induction of specific neutralizing antibodies for potentia l immunotherapentic applications for the prevention of HRV infection. (C) 2 001 Academic Press.