Departments of Pathobiology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
J Virol Methods. 2021 Aug;294:114172. doi: 10.1016/j.jviromet.2021.114172. Epub 2021 Apr 26.
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.
为了展示和描绘该序列的启动子功能，将 EGFP 表达质粒 pEGFP-N1 中的 CMV 早期启动子替换为禽腺病毒 9（FAdV-9）基因组的非常左端（ntds73-574）。表达取代 ORF1 和 ORF2 的 EGFP ORF 表明，该天然启动子能够驱动下游外源基因的表达。用包含来自安大略省禽脑脊髓炎病毒（AEV）的 493bp IRES 的双顺反子盒替换 ORF1 和 ORF2，该 IRES 将 EGFP ORF 和 mCherry ORF 分开，允许重组 FAdV 表达两个 ORF。这些结果为基于天然 FAdV-9 启动子和禽病毒 IRES 的多价疫苗开发提供了另一个平台。
