The genome of the multiple-embedded nuclear polyhedrosis virus (MNPV) of (LdMNPV) was partially characterized by restriction endonuclease analysis and a physical map was constructed using cosmid cloning and Southern cross blot hybridization. Using HI, II, RI and dIII, the size of the genome was estimated to be 88.5 × 10 or 134.04 kbp. LdMNPV DNA was also analysed using methylation-sensitive restriction enzymes. The resulting restriction profiles suggested that extensive methylation did not occur at the nucleotide sequence recognized by II and I. A HI restriction map was constructed by comparing overlapping HI fragments between cosmid clones containing partial digests of viral DNA. The positions of the II, RI and dIII sites were determined by Southern cross blot hybridizations and aligned to the HI restriction map. At least four homologous regions were identified by cross blot hybridizations of II-digested LdMNPV DNA and such regions were found to be interspersed along the genome in a fashion similar to that reported for other baculoviruses. Using recombinant plasmids containing the dIII-V fragment of MNPV to probe Southern blots of LdMNPV DNA, the restriction fragment(s) that contain the polyhedrin gene were identified. Based on these findings the map was oriented with the polyhedrin gene of LdMNPV as the zero point.

Keyword(s): baculovirus , L. dispar and polyhedrin

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