[Sequencing of polyhedrin gene of Leucania separata multiple capsid nuclear polyhedrosis virus by silver sequencing...

The genome of LsNPV was digested with EcoRV and cloned into pBluescript. With the partial polyhedrin gene of Autographa californica as a probe, positive recombinants were screened by colony in situ hybridization, further characterized by sequencing, and designated as pLsEV and pLsPH5. The intact 741 bp polyhedrin gene of LsNPV is located on the two recombinants revealed by Silver Sequencing System, and shares 80.0% and 97.0% nucleotide identity of the coding sequence, 87.8% and 97.5% amino acid identity with AcMNPV and MbMNPV respectively. Glu is the richest amino acid, and Gln and Trp the poorest amino acid in the polyhedrin of LsNPV. The codons ended with pyrimidine were higher in the codon usage. There was a signal-peptide-like hydrophobic region of 26 amino acids in the N end of polyhedrin.