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This dissertation, The Analysis of 5' and 3' Untranslated Regions (UTRS) of Influenza A Virus by Shuk-fan, Ng, 吳淑芬, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of the thesis entitled The analysis of 5' and 3' untranslated regions (UTRs) of influenza A virus Submitted by Ng Shuk Fan for the Degree of Master of Philosophy at The University of Hong Kong in August 2005 Influenza A virus is an enveloped virus that consists of 8 segments of virion RNA (vRNA) of negative polarities. Each of eight vRNA segments has a promoter composed of sequences derived from both ends. The promoter sequence at each end includes both the universal conserved region and the segment specific region. Previous studies on the universal conserved sequences have shown that these regions are important in regulating viral transcription, translation, replication and RNA packaging. However, the studies on the segment specific region are limited. In order to understand the functions of the segment specific regions of vRNA on the viral life-cycle, a reverse genetics system was employed to generate a recombinant virus with mutations in the segment specific regions of nonstructural (NS) protein vRNAs. The generated viral mutant, NSNA, contains a NS coding sequence flanked by neuraminidase (NA) promoter sequences. In addition, a pair of mutants, All-U and NSNA-U were generated which are genetically identical to the wild type and th NSNA viruses, except that there is a C to U mutation at the 4 residue of the 3' ends of three polymerases vRNA segments. The growth characteristics of the recombinant viruses were analyzed by plaque assays, real-time PCR, western blotting, RNA gel electrophoresis and electron microscopy. The results demonstrated that the recombinant viruses were attenuated in MDCK cells. Cells infected with the NSNA and NSNA-U mutants were found to have elevated NS and NA vRNA expression, but the mutations had no effect on the PB2 vRNA level. The NS vRNA levels in viral particles were increased. In contrast, their mRNA levels in infected cells and protein expressions were not affected by the mutations. These data indicated that the segment specific region may play a role in regulating the viral replication and virion assembly. (293 words) DOI: 10.5353/th_b3572322 Subjects: Genetic translationInfluenza A virus - Genetic aspects

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