With respect to proposed novel ORFs identified by in silico analysis, in most cases, we found no evidence for transcript expression. Of those that gave positive hybridisation signals, most might be explained by overlapping transcripts from genes in the same region and coding in the same direction. Other novel ORFs lie within regions of the genome now considered to be non-coding, but where transcripts have previously been reported, while the remainder may represent genuine coding ORFs. The lack of signal for previously described ORFs that are now considered non-protein coding confirms their status as discounted genes. In order to test the microarray system, the temporal expression kinetics of selected virus genes were investigated by alternative methods including; real-time PCR, and northern blots to check the identity of specific transcripts, and where antibodies were available, western immunoblotting to confirm the expression kinetics of specific proteins. The expression kinetics obtained for specific genes both differentially and non-differentially expressed by these various methods were entirely consistent with those obtained for the same genes with the HCMV microarray. It was concluded that the Merlin microarray system was a valid and reliable research tool for the investigation of HCMV gene expression.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:519435 |
| Date | January 2007 |
| Creators | Towler, James Charles |
| Publisher | University of Glasgow |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://theses.gla.ac.uk/42/ |
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