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Analysis of the fowlpox virus homologue of mammalian PC-1 glycoprotein

Anwar, Mohammad Arif; (1999) Analysis of the fowlpox virus homologue of mammalian PC-1 glycoprotein. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

Fowlpox virus (FWPV) is the prototypic member of the genus Avipoxviridae. Although no longer a commercial threat to livestock, in recent years it has been extensively used for expression of foreign antigens, as recombinant vaccines for avian and non-avian targets. Despite this attention, FWPV remains poorly characterised. Sequence analysis of a 6.5kbp region near the left inverted terminal repeat of the FWPV genome had revealed five major ORFs not previously observed in any other virus. One of these ORFs (FP-PC1), exhibited 39% amino acid identity to mammalian PC1 glycoprotein, initially described as an antigen of terminally differentiated B-cells. The full length FP-PC1 gene was cloned directly from the FWPV genome using specific primers. Using the transient dominant selection technique, the resultant transfer vector was used to construct three recombinant FWPV, with different deletions in the FP-PC1 ORF. The FP-PC1 gene was shown to be non-essential for FWPV replication in vitro, as FWPV with a full gene deletion in the FP-PC1 ORF was readily isolated. Further characterisation of the mutant FWPV indicated that FP- PC1 did not affect FWPV replication in vitro, although a difference in plaque size was observed. A possible involvement of FP-PC1 in the nucleotide salvage pathway was investigated by comparing wt and mutant FWPV growth in nucleotide deficient media. But no difference was observed. Thus further investigation needs to be conducted to investigate a potential role for FP-PC1 as a potential scavenger of nucleotides. Both the full length and extracellular FP-PC1 domains were expressed in an in vitro cell free system using rabbit reticulocyte lysates. They were also overexpressed in FWPV infected cells, co-infected with a recombinant FWPV expressing T7 polymerase. Using data from both these systems, full length FP-PC1 was shown to be an N-linked glycosylated integral membrane protein with a type II configuration.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Analysis of the fowlpox virus homologue of mammalian PC-1 glycoprotein
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Biological sciences; Open reading frames
URI: https://discovery.ucl.ac.uk/id/eprint/10120756
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