Characterization of a fowl adenovirus 9 (FAdV-9) early promoter and its application in generating dual expression FAdV-9s

Yanlong Pei; Peter J Krell; Leonardo Susta; Éva Nagy

doi:10.1016/j.jviromet.2021.114172

Characterization of a fowl adenovirus 9 (FAdV-9) early promoter and its application in generating dual expression FAdV-9s

J Virol Methods. 2021 Aug:294:114172. doi: 10.1016/j.jviromet.2021.114172. Epub 2021 Apr 26.

Authors

Yanlong Pei¹, Peter J Krell², Leonardo Susta¹, Éva Nagy³

Affiliations

¹ Departments of Pathobiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
² Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
³ Departments of Pathobiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada. Electronic address: enagy@uoguelph.ca.

PMID: 33915232
DOI: 10.1016/j.jviromet.2021.114172

Abstract

The CMV immediate early promoter from the EGFP expression plasmid pEGFP-N1 was replaced with the very left end of the fowl adenovirus 9 (FAdV-9) genome (ntds 73-574) to demonstrate and delineate the promoter function of this sequence. Expression of an EGFP ORF which replaced ORF1 and ORF2 demonstrated that the native promoter can drive down stream foreign gene expression. Replacement of ORF1 and ORF2 with a bicistronic cassette, incorporating a 493 bp IRES from an Ontario strain of avian encephalomyelitis virus (AEV) separating an EGFP ORF and mCherry ORF allowed for expression of both ORFs from a recombinant FAdV. These results provide an additional platform for multivalent vaccines development based on a native FAdV-9 promoter and an avian virus IRES.

Keywords: Avian encephalomyelitis virus IRES; Bicistronic cassette; FAdV-9; Native promoter.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenoviridae Infections*
Animals
Aviadenovirus* / genetics
Chickens
Fowl adenovirus A* / genetics
Gene Expression
Open Reading Frames
Plasmids
Poultry Diseases*
Promoter Regions, Genetic