Cloning sequencing and expression of the gene encoding the VP2 protein of the human group B rotavirus, ADRV.
Adult diarrheal rotavirus, ADRV, is a noncultivable human group B rotavirus. A complete cDNA copy of ADRV gene segment 2 has been cloned and sequenced. Gene segment 2 contains 2844 bases and encodes one long open reading frame beginning at base 14 and terminating at base 2812. Gene 2 encodes a protein containing 933 amino acids with a calculated molecular weight of 105.6 kDa and a pl of 5.5. The gene 2 polypeptide contains significant homology with the VP2 protein which comprises the core of group A rotavirus strains. The gene 2 protein has been expressed in a rabbit reticulocyte lysate in vitro and is identical in molecular mass with a protein previously demonstrated to be present on iodinated EDTA-treated virions and from in vitro translations of total ADRV mRNA. A recombinant baculovirus containing gene segment 2 has been constructed and used to express the encoded VP2 equivalent protein. The expressed VP2 protein is immunoprecipitable by hyperimmune anti-ADRV serum, porcine group B infection serum, and human convalescent serum but not by hyperimmune serum to group A rotavirus. Our results suggest that ADRV gene segment 2 encodes the VP2 protein equivalent to group A rotavirus strains present in the core of the group B virion.
Mackow
E R
ER
Northport VA Medical Center, New York 11768.
Fay
M E
ME
Shaw
R
R
Tao
H
H
Chen
G
G
D000595:Amino Acid Sequence; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D036022:Capsid Proteins; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000032:analysis; D012401:Rotavirus / Q000235:genetics* / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017423:Sequence Analysis, RNA; D017386:Sequence Homology, Amino Acid
D036022:Capsid Proteins; D011994:Recombinant Proteins; C442545:VP2 protein, Rotavirus