Deletion of the AcMNPV core gene ac109 results in budded virions that are non-infectious.
Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac109 is a core gene and its function in the virus life cycle is unknown. To determine its role in the baculovirus life cycle, we used the AcMNPV bacmid system to generate an ac109 deletion virus (vAc(109KO)). Fluorescence and light microscopy showed that transfection of vAc(109KO) results in a single-cell infection phenotype. Viral DNA replication is unaffected and the development of occlusion bodies in vAc(109KO)-transfected cells evidenced progression to the very late phases of viral infection. Western blot and confocal immunofluorescence analysis showed that AC109 is expressed in the cytoplasm and nucleus throughout infection. In addition, AC109 is a structural protein as it was detected in both budded virus (BV) and occlusion derived virus in both the envelope and nucleocapsid fractions. Titration assays by qPCR and TCID(50) showed that vAc(109KO) produced BV but the virions are non-infectious. The vAc(109KO) BV were indistinguishable from the BV of repaired and wild type control viruses as determined by negative staining and electron microscopy.
Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, Box 5000, Summerland BCV0H1Z0, Canada.
Minggang
M
Fang
Yingchao
Y
Nie
David A
DA
Theilmann
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000096:biosynthesis; D017353:Gene Deletion; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology; D018411:Spodoptera / Q000821:virology*; D014758:Viral Core Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics*; D014779:Virus Replication
D004279:DNA, Viral; D014758:Viral Core Proteins