Location: Endemic Poultry Viral Diseases ResearchTitle: Recombineering of the infectious laryngotracheitis genome using synthetic genomics assembly
|GARCIA, MARICARMEN - University Of Georgia|
|FUCHS, WALTER - Friedrich-Loeffler-institut|
|RIBLET, SYLVA - University Of Georgia|
|VOLKENING, JEREMY - Base2bio|
Submitted to: Meeting Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 7/13/2018
Publication Date: 7/13/2018
Citation: Spatz, S.J., Garcia, M., Fuchs, W., Ross, T.A., Riblet, S., Kim, T.N., Miller, P.J., Volkening, J., Afonso, C.L. 2018. Recombineering of the infectious laryngotracheitis genome using synthetic genomics assembly. Proceedings of the American Association of Avian pathologists Annual Meeting, Denver, Colorado, July 13-17, 2018.
Technical Abstract: The study of gene function of the herpesvirus, infectious laryngotracheitis virus (ILTV) has been difficult because of the lack of an infectious clone. To overcome this, cosmid and yeast centromere plasmid libraries were generated and the nucleotide sequences of individual clones determined. Viable virus could be reconstituted in vitro using a set of overlapping clones representative of the full ILTV genome. To demonstrate the usefulness of this system, we generated mutant viruses containing deletions and insertions. Chicken kidney (CK) cells infected with these mutants exhibited growth kinetics similar to that of the wild type parental virus. To determine whether the reconstituted viruses retained their virulent phenotypes, specific pathogen free chickens were inoculated with the viruses and succumbed to clinical disease to similar degrees as birds inoculated with wild type virus. Overall this study demonstrated that viable virus could be reconstituted from a collection of recombinant clones and a unique locus was identified to which foreign genes could be inserted into the ILTV genome without affecting in vitro and in vivo growth properties.