ANALYSIS OF VARIATION IN INFLUENZA A VIRUS GENES.
Item
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Title
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ANALYSIS OF VARIATION IN INFLUENZA A VIRUS GENES.
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Identifier
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AAI8203264
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identifier
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8203264
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Creator
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BAEZ, MELVYN.
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Contributor
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Dr. Peter Palese
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Date
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1981
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Language
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English
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Publisher
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City University of New York.
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Subject
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Engineering, Biomedical
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Abstract
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The genes of several influenza A viruses were examined by using various biochemical techniques and molecular cloning.;The genetic compositions of eleven high yielding recombinants of influenza virus were determined by polyacrylamide gel electrophoresis of the ('32)P-labeled RNAs obtained from the recombinants and their parents. The enhanced growth capacity of the recombinants, constructed for use in vaccine production, was associated with the presence of genes derived from the high yielding laboratory strain A/PR/8/34. In particular, all of the examined high yielding recombinants derived the M gene from the A/PR/8/34 strain.;Two dimensional electrophoresis of ribonuclease T1 resistant oligonucleotides was used to estimate the extent of genetic variation among five different influenza viruses isolated during the beginning, middle and end of the H2N2 subtype period (1957-1968). It was demonstrated that genetic variation occurred among all the corresponding RNAs of these influenza viruses and that the extent of variation was similar for all genes. It is suggested that the maximum mutational changes of 3 or 4%, that were estimated for all the genes of the H2N2 viruses, may represent the extent of genetic variation that occurred during the eleven year interval which separates the isolation of the earliest and latest H2N2 strain examined in this study.;The NS genes of the human influenza virus, A/PR/8/34, and the avian influenza virus, A/duck/Alberta/60/76, were cloned into pBR322. The complete nucleotide sequences of these two NS genes were determined and a comparison of these sequences showed a 27.3% difference. In contrast, the differences among the NS gene of the A/PR/8/34 virus and the previously sequenced NS genes of the avian virus A/FPV/Rostock/34 and the human virus A/Udorn/72 were fewer in number and ranged from 8-11%. The extensive sequence similarity among these latter three strains does not support the suggestion of species specific homology groups among the NS genes of avian and human influenza viruses. The NS genes of the A/PR/8/34, A/Udorn/72 and A/FPV/Rostock/34 strains were shown to contain open reading frames in their virion (-) RNAs which potentially encode polypeptides of 167 and 216 amino acids. Although the NS segment of the A/duck/Alberta/60/76 virus lacks an agnogene sequence in its negative strand sequence, it conserves the overlapping NS1 and NS2 gene arrangement identified in the other NS RNA segments. A comparison of the deduced amino acid sequences of the A/duck/Alberta/60/76 and A/PR/8/34 virus NS genes showed that the NS2 polypeptides are more highly conserved (18.2%) than the NS1 polypeptides (33.0% difference). Surprisingly, the NS1 polypeptides of the two strains showed this high degree of variation over their entire lengths, including the amino acids encoded by the conserved RNA domain of the NS1 and NS2 gene overlap.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biomedical Sciences
