Expression of fowl adenovirus type 10 antigens in Escherichia coli.
In an attempt to construct a genetic map of the fowl adenovirus (FAV) and to determine which viral proteins are natural immunogens in chickens and hence may be relevant to protective immunity we have constructed an expression library of FAV type 10 DNA. The genomic DNA was partially digested with the restriction endonuclease Sau3A, and this DNA was inserted into the 3' terminal end of the beta-galactosidase gene in a plasmid vector. To date, approximately 600 clones have been identified that express FAV type 10 antigens as determined by immunological screening with rabbit antisera to purified virus, including one that has amino acid homology with the 100 kDa protein of human adenovirus type 5. These antigen positive clones were found to contain DNA from FAV type 10 genome as determined by hybridisation to FAV DNA. These clones will allow the further characterisation of FAV and possibly the identification of potential vaccine molecules.
Sheppard
M
M
CSIRO, Animal Health Research Laboratory, Parkville, Victoria, Australia.
Werner
W
W
D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics*; D000259:Aviadenovirus / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D002645:Chickens; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D015723:Gene Library; D005822:Genetic Vectors; D007118:Immunoassay; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015183:Restriction Mapping; D013047:Specific Pathogen-Free Organisms
D000956:Antigens, Viral; D004279:DNA, Viral