Identification of non-essential loci within the Meleagrid herpesvirus 1 genome

Hall, Robyn N., Meers, Joanne, Fowler, Elizabeth V. and Mahony, Timothy J. (2015) Identification of non-essential loci within the Meleagrid herpesvirus 1 genome. Virology Journal, 12 1: 130.1-130.12. doi:10.1186/s12985-015-0362-9

Author Hall, Robyn N.
Meers, Joanne
Fowler, Elizabeth V.
Mahony, Timothy J.
Title Identification of non-essential loci within the Meleagrid herpesvirus 1 genome
Formatted title
Identification of non-essential loci within the Meleagrid herpesvirus 1 genome
Journal name Virology Journal   Check publisher's open access policy
ISSN 1743-422X
Publication date 2015-08-27
Sub-type Article (original research)
DOI 10.1186/s12985-015-0362-9
Open Access Status DOI
Volume 12
Issue 1
Start page 130.1
End page 130.12
Total pages 12
Place of publication London, United Kingdom
Publisher BioMed Central
Collection year 2016
Language eng
Formatted abstract
Background:  Meleagrid herpesvirus 1 (MeHV-1) infectious bacterial artificial chromosomes (iBACs) are ideal vectors for the development of recombinant vaccines for the poultry industry. However, the full potential of iBACS as vectors can only be realised after thorough genetic characterisation, including identification of those genetic locations that are non-essential for virus replication. Generally, transposition has proven to be a highly effective strategy for rapid and efficient mutagenesis of iBAC clones. The current study describes the characterisation of 34 MeHV-1 mutants containing transposon insertions within the pMeHV1-C18 iBAC genome.

Methods:  Tn5 and MuA transposition methods were used to generate a library of 76 MeHV-1 insertion mutants. The capacity of each mutant to facilitate the recovery of infectious MeHV-1 was determined by the transfection of clone DNA into chicken embryo fibroblasts.

Results:  Attempts to recover infectious virus from the modified clones identified 14 genetic locations that were essential for MeHV-1 replication in cell culture. Infectious MeHV-1 was recovered from the remaining 14 intragenic insertion mutants and six intergenic insertion mutants, suggesting that the respective insertion locations are non-essential for MeHV-1 replication in cell culture.

Conclusions:  The essential and non-essential designations for those MeHV-1 genes characterised in this study were generally in agreement with previous reports for other herpesviruses homologues. However, the requirement for the mardivirus-specific genes LORF4A and LORF5 are reported for the first time. These findings will help direct future work on the development of recombinant poultry vaccines using MeHV-1 as a vector by identifying potential transgene insertion sites within the viral genome.
Keyword Meleagrid herpesvirus 1
Infectious bacterial artificial chromosomes
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Alliance for Agriculture and Food Innovation
Official 2016 Collection
School of Veterinary Science Publications
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