The UL47 gene of avian infectious laryngotracheitis virus is not essential for in vitro replication but is relevant for virulence in chickens.
The genome of infectious laryngotracheitis virus (ILTV) exhibits several differences from those of other avian and mammalian alphaherpesviruses. One of them is the translocation of the conserved UL47 gene from the unique long (UL) to the unique short (US) genome region, where UL47 is inserted upstream of the US4 gene homologue. As in other alphaherpesviruses, UL47 encodes a major tegument protein of ILTV particles, whereas the US4 gene product is a non-structural glycoprotein, gG, which is secreted from infected cells. For functional characterization, an ILTV recombinant was isolated in which US4 together with the 3'-terminal part of UL47 was replaced by a reporter gene cassette encoding green fluorescent protein. From this virus, UL47 and US4 single-gene deletion mutants without foreign sequences were derived and virus revertants were also generated. In vitro studies revealed that both genes were non-essential for ILTV replication in cultured cells. Whereas US4-negative ILTV exhibited no detectable growth defects, maximum virus titres of the double deletion mutant and of UL47-negative ILTV were reduced about 10-fold compared with those of wild-type virus and rescued virus. Experimental infection of chickens demonstrated that UL47-negative ILTV was significantly attenuated in vivo and was shed in reduced amounts, whereas wild-type and rescued viruses caused severe disease and high mortality rates. As all immunized animals were protected against subsequent challenge infection with virulent ILTV, the UL47 deletion mutant might be suitable as a live-virus vaccine.
Helferich
Dorothee
D
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
Veits
Jutta
J
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
Teifke
Jens P
JP
Institute of Infectology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
Mettenleiter
Thomas C
TC
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
Fuchs
Walter
W
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D017353:Gene Deletion; D017930:Genes, Reporter; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006566:Herpesviridae Infections / Q000473:pathology / Q000503:physiopathology / Q000662:veterinary / Q000821:virology; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D006651:Histocytochemistry; D008168:Lung / Q000473:pathology; D011201:Poultry Diseases / Q000821:virology; D011995:Recombination, Genetic; D014132:Trachea / Q000473:pathology; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics* / Q000502:physiology; D014774:Virulence / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
D014764:Viral Proteins; D049452:Green Fluorescent Proteins