Isolation, Complementation and Partial Characterization of Temperature-sensitive Mutants of the Baculovirus Hyphantria Cunea Nuclear Polyhedrosis Virus
Twelve temperature-sensitive mutants were isolated from Hyphantria cunea nuclear polyhedrosis virus grown in Spodoptera frugiperda cells and were sorted into four groups by their properties in plaque assays at the non-permissive temperature (32 °C). The phenotypes of the four groups were as follows: (i) failure to make polyhedra, (ii) few polyhedra formed, (iii) reduced plaquing efficiency, (iv) small plaque size. Ten mutants had reduced plaque size and polyhedra formation at 32 °C. One mutant formed plaques without polyhedra, had a reduced infectious virus titre at 32 °C and also showed a defect in late gene function. Two mutants formed small plaques with few polyhedra and were temperature-sensitive with respect to production of extracellular non-occluded virions at 32 °C. Other phenotypes were also distinguished. The formation of polyhedra by all the mutants was 2 to 4 h faster at 32 °C than at 25 °C. After temperature shift-up from 25 °C to 32 °C at 12 h post-infection polyhedron formation was still 2 to 4 h faster. Complementation analyses based on polyhedron formation in double infections at 32 °C distinguished four complementation groups.
BROWNM., FAULKNERP.1980; A partial genetic map of the baculovirus, Autographa californica nuclear polyhedrosis virus, based on recombination studies with ts mutants. Journal of General Virology 48:247–251
BROWNM., CRAWFORDA. M., FAULKNERP.1979; Genetic analysis of a baculovirus Autographa californica nuclear polyhedrosis virus. 1. Isolation of temperature-sensitive mutants and assortment into complementation groups. Journal of Virology 31:190–198
CHOEY. K., BYUNS. M., LEEH. H., CHUNGT. W.1986; A nuclear polyhedrosis virus of Hyphantria cunea: replication in vitro. Biotechnology Letters 8:853–858
DUNCANR., FAULKNERP.1982; Bromodeoxyuridine-induced mutants of Autographa californica nuclear polyhedrosis virus defective in occlusion body formation. Journal of General Virology 62:369–373
GORDONJ. D., CARSTENSE. B.1984; Phenotypic characterization and physical mapping of a temperature-sensitive mutant of Autographa californica nuclear polyhedrosis virus defective in DNA synthesis. Virology 138:69–81
HINKW. F., STRAUSSE.1976; Replication and passage of alfalfa looper NPV plaque variants in cloned cell cultures and larval stages of four host species. Journal of Invertebrate Pathology 27:49–55
KNUDSOND. L., TINSLEYT. W.1974; Replication of a nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda: purification, assay of infectivity, and growth characteristics of the virus. Journal of Virology 14:934–944
KNUDSOND. L., HARRAPK. A.1976; Replication of a nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda: microscopy study of the sequence of events of the virus infection. Journal of Virology 17:254–268
LEEH. H.1987 Replication and cloning of Hyphantria cunea nuclear polyhedrosis virus in Spodoptera frugiperda cell line. Hanguk Journal of Genetic Engineering21–6 Seoul: Kon Kuk University;
LEEH. H., MILLERL. K.1979; Isolation, complementation, and initial characterization of temperature-sensitive mutants of the baculovirus Autographa californica nuclear polyhedrosis virus. Journal of Virology 31:240–252
MILLERL. K.1981; Construction of a genetic map of the baculovirus Autographa californica nuclear polyhedrosis virus by marker rescue of temperature-sensitive mutants. Journal of Virology 39:973–980
MINE. S., LEEH. H.1982; Electron microscopic visualization of temperature-sensitivity of Autographa californica NPV ts-B1074 mutant. Journal of the Korean Society of Virology 12:49–54
OHC. K., LEEH. H.1987; Electron microscope studies of infection and multiplication of Autographa californica NPV clone L-1 and ts-B1074. 1. Infection and multiplication of A. californica NPV L-1. Journal of the Korean Society of Virology 17:61–72
POTTERK. N., FAULKNERP., MACKINNONE. A.1976; Strain selection during serial undiluted passage of Trichoplusia ni nuclear polyhedrosis virus. Journal of Virology 18:1040–1050
ROHELD. Z., COCHRANM. A., FAULKNERP.1983; Characterization of two abundant mRNAs of Autographa californica nuclear polyhedrosis virus present late in infection. Virology 124:357–365
SMITHG. E., SUMMERSM. D.1979; Restriction maps of five Autographa californica MNPV variants, Trichoplusia ni MNPV, and Galleria mellonella MNPV DNAs with endonucleases SmaI, KpnI, BamHI, Sacl, XhoI, and EcoRI. Journal of Virology 30:828–838
SUMMERSM. D., ARNOTTH. J.1969; Ultrastructural studies on inclusion formation and virus occlusion in nuclear polyhedrosis and granulosis virus-infected cells of Trichoplusia ni (Hübner). Journal of Ultrastructure Research 28:462–80
VAUGHNJ. L., GOODWINR. H., TOMKINSG. J., MCCAWLEYP.1977; The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13:213–217
VLAKI. M.1981; Mapping of BamHI and SmaI DNA restriction sites on the genome of the nuclear polyhedrosis virus of the alfalfa looper, Autographa californica. Journal of Invertebrate Pathology 36:409–414
VLAKJ. M., SMITHG. E., SUMMERSM. D.1981; Hybridization selection and in vitro translation of Autographa californica nuclear polyhedrosis virus mRNA. Journal of Virology 40:762–771
VOLKMANL. E., SUMMERSM. D.1977; Autographa californica NPV: comparative infectivity of the occluded, alkali-liberated, and nonoccluded forms. Journal of Invertebrate Pathology 30:102–103
VOLKMANL. E., SUMMERSM. D., HSIEHC. H.1976; Occluded and nonoccluded nuclear polyhedrosis virus grown in Trichoplusia ni: comparative neutralization, comparative infectivity, and in vitro growth studies. Journal of Virology 19:820–832
Isolation, Complementation and Partial Characterization of Temperature-sensitive Mutants of the Baculovirus Hyphantria Cunea Nuclear Polyhedrosis Virus