Bovine respiratory syncytial virus nucleocapsid protein: mRNA sequence analysis and expression from recombinant vaccinia virus vectors

• Published in: Journal of general virology Vol. 73; no. 4; pp. 999 - 1003
• Main Authors: Amann, V.L, Lerch, R.A, Anderson, K, Wertz, G.W
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.04.1992; Society for General Microbiology
• Subjects: Amino Acid Sequence; amino acid sequences; Animals; Base Sequence; Biological and medical sciences; Bovine respiratory syncytial virus; Capsid - biosynthesis; Capsid - genetics; Cattle; Cells, Cultured; clones; Cloning, Molecular; DNA; DNA, Single-Stranded - genetics; Fundamental and applied biological sciences. Psychology; Genetic Vectors; Genetics; messenger RNA; Microbiology; Molecular Sequence Data; nucleotide sequences; Recombinant Proteins - biosynthesis; respiratory syncytial virus; Respiratory Syncytial Viruses - genetics; RNA, Messenger - genetics; Sequence Homology, Nucleic Acid; Transfection; Vaccinia virus - genetics; Viral Core Proteins - biosynthesis; Viral Core Proteins - genetics; Virology; Chordopoxvirinae; Nucleotide sequence; Recombinant virus; Orthopoxvirus; Sequence alignment; Paramyxoviridae; Virus; Proteins; Vaccinia virus; Messenger RNA; Poxviridae; Nucleocapside; Pneumovirus; Bovine pneumovirus; Comparative study
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-73-4-999

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 Mr of 42.6 K. 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.