CHARACTERIZATION AND PARTIAL CLONING OF THE GENOMIC DNA OF A BACULOVIRUS FROM PENAEUS-JAPONICUS (PJNOB=BMNV)

Many particles of a non-occluded baculovirus with a membrane 70 x 330 nm in size were observed in the nuclei of the mid-gut glands of larval Kuruma shrimp (Penaeus japonicus) with mid-gut necrosis using an elec tron microscope. Nucleotides of the baculovirus were isolated from the diseased larvae in the presence of Triton X-100. The nucleocapsid was a bacilliform virion, approximately 50 nm in width and 260 nm in leng th, From the purified preparations of nucleocapsids of the virus, two major proteins with molecular weights of 35 and 14 kDa were identified by SDS-polyacrylamide gel electrophoretic analysis, The viral genomic DNA extracted from the nucleocapsids was digested with BamHI and Sau3 AI, but not with EcoRI, PstI, XhoI and Sail. The molecular weight of t he DNA was estimated to be approximately 85.1 X 10(6) from the agarose gel electrophoretic pattern of the BamHI fragments, A part of the Bam HI fragments was cloned, labeled with P-32-CTP and used for Southern b lot analysis as a probe. The fragment: hybridized to DNA isolated from the diseased larvae, but not to DNA isolated from healthy larvae.