Bovine respiratory syncytial virus nucleocapsid protein: mRNA sequence analysis and expression from recombinant vaccinia virus vectors.
The nucleotide sequence of the mRNA encoding the nucleocapsid (N) protein of bovine respiratory syncytial (BRS) virus, strain 391-2, was determined. Recombinant vectors containing a cDNA of the complete N gene were constructed, and expression of the N protein in eukaryotic cells was demonstrated using two different vector systems. The BRS virus N mRNA was 1197 nucleotides in length, exclusive of poly(A), and had a single major open reading frame that encoded a polypeptide of 391 amino acids with a calculated M(r) of 42.6K. The nucleotide and amino acid sequences of the BRS virus N gene were compared to those of human respiratory syncytial (HRS) virus strains A2 and 18537, and to BRS virus strain A51908. The level of nucleic acid identity between the N mRNA of BRS virus 391-2 and both HRS virus subtypes was 80 to 81%, whereas the identity between the two BRS virus strains was 97%. A 93 to 94% level of identity was observed between the deduced amino acid sequences of the N protein of BRS virus 391-2 and the corresponding sequences of the two HRS virus strains. The two BRS virus N proteins differed in amino acid sequence at only three positions. Recombinant BRS virus N protein was expressed using two different vector systems: in cells from a plasmid using the vaccinia virus/T7 polymerase expression system or from a recombinant vaccinia virus. N proteins synthesized by the two vector systems migrated with an electrophoretic mobility identical to that of authentic BRS virus N protein, and were precipitated by anti-BRS virus antibodies.
Amann
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VL
Department of Microbiology, University of Alabama, Birmingham 35294.
Lerch
R A
RA
Anderson
K
K
Wertz
G W
GW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics*; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012136:Respiratory Syncytial Viruses / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics*
D004277:DNA, Single-Stranded; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014758:Viral Core Proteins