Base sequence differences among the ribonucleic acids of influenza virus.
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Biomedical subjects
Publications and source records attributed to J Content.
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Ribonucleic acid extracted from influenza virus was labeled at the 3' termini with (3)H and analyzed by polyacrylamide gel electrophoresis. Influenza virus was found to contain a minimum of seven and possibly as many as 10 polynucleotide chains, most of which appear to terminate at the 3' end in uridine.
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The proteins of influenza (WSN) and Sendai virus have been separated by polyacrylamide gel electrophoresis into five components. In both cases, three of these components were shown to be glycoproteins containing fucose, galactose, and glucosamine. Two protein components of each virus were probably free from these sugar residues, including the structural unit of the viral ribonucleoprotein (molecular weight of about 60,000 daltons).
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Human fibroblasts were induced for interferon synthesis, and the mRNA coding for interferon was partially purified. On this basis, double-stranded DNA copies were synthesized enzymatically and were inserted into cloning vehicles. A large collection of colonies containing chimeric plasmids was obtained. An RNA selection method was used for the identification of several individual clones containing the human fibroblast interferon (IFN-beta) cDNA. From the nucleotide sequence of the cloned structural gene, the primary structure of the mRNA and, hence, of the protein itself was deduced. IFN-beta is a polypeptide 166 amino acids long and contains a single site for N-glycosylation; IFM-beta is normally made as a preinterferon containing a signal sequence that is 21 amino acids long. The interferon gene was inserted into an expression plasmid under thermoinducible control of the phage lambda PL promoter. this allowed the synthesis of polypeptides in the bacteria, which for all physicochemical, biological, and immunologic properties tested closely resembled authentic IFN-beta.