Construction of recombinant avian infectious laryngotracheitis virus expressing the beta-galactosidase gene and DNA sequencing of the insertion region.
Avian infectious laryngotracheitis virus (ILTV), a herpesvirus, is a highly contagious pathogen that causes an upper respiratory tract infection in chickens. It is one of the major problems in the poultry industry worldwide. Current vaccines are not satisfactory due to the induction of latent infection. Here we describe a system for the construction of recombinant ILTV. A 4-kbp ILTV EcoRI DNA fragment was cloned into plasmid pUC13 and sequenced. Computer prediction revealed two potential open reading frames with 216 and 259 amino acid residues, respectively. The 259-residue polypeptide was serine-rich. The beta-galactosidase (beta-gal) gene of E. coli was cloned into the XhoI/Bg/II site of this DNA fragment, integrated into the ILTV genome via homologous recombination, expressed under the control of the immediate-early cytomegalovirus promoter, and caused the formation of blue plaques in the presence of X-gal. The insertion of a foreign gene into the ILTV genome and the successful expression of the incorporated gene demonstrated the potential for the construction of attenuated recombinant ILTV vaccines and the development of ILTV as vectors for polyvalent vaccines against avian upper respiratory tract infections.
Guo
P
P
Department of Veterinary Pathobiology, Purdue University, West Lafayette, Indiana 47907-1243.
Scholz
E
E
Maloney
B
B
Welniak
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers / Q000737:chemistry; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics; D005822:Genetic Vectors; D001352:Herpesvirus 1, Gallid / Q000235:genetics* / Q000254:growth & development; D066298:In Vitro Techniques; D008114:Liver Neoplasms, Experimental; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured; D014779:Virus Replication; D001616:beta-Galactosidase / Q000235:genetics*
D017931:DNA Primers; D004274:DNA, Recombinant; D004279:DNA, Viral; D001616:beta-Galactosidase