Expression of bovine rotavirus neutralization antigen in Escherichia coli.
A 646 bp fragment derived from a full length cDNA clone of genomic segment 9 of bovine rotavirus (NCDV strain) was inserted into Escherichia coli expression plasmid pEX1. The fragment encodes amino acids 50 to 265 of the major vital neutralization antigen VP7, a 326 amino acid long outer shell glycoprotein. Several transformed bacterial clones were isolated in which the recombinant plasmid directed the synthesis of a cro-beta-galactosidase-VP7 fusion protein that was recognized by rabbit polyclonal antibodies against NCDV rotavirus. Sera from rabbits immunized with the fusion protein specifically reacted with VP7 among NCDV virion polypeptides. The chimeric polypeptide was also specifically recognized by two monoclonal antibodies against UK strain rotavirus VP7 that exhibited virus-neutralizing activity. However, immune sera to the chimeric polypeptide showed no neutralizing activity against bovine rotavirus. These results are discussed in view of a recent report that a fusion VP7-beta-galactosidase polypeptide comprising 35 more amino acids at the carboxy terminus was able to induce neutralizing antibodies in mice to simian rotavirus SA11.
Francavilla
M
M
Istituto di Genetica Biochimica ed Evoluzionistica CNR, Pavia, Italy.
Miranda
P
P
Di Matteo
A
A
Sarasini
A
A
Gerna
G
G
Milanesi
G
G
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D008970:Molecular Weight; D009500:Neutralization Tests; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012401:Rotavirus / Q000235:genetics*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D004247:DNA