›› 2012, Vol. 55 ›› Issue (4): 403-411.doi:

• RESEARCH PAPERS • Previous Articles     Next Articles

Molecular cloning and cellular localization of Opdi108 from Ophiusa disjungens nucleopolyhedrovirus

CHANG Run-Lei, LIU Li, WEI Chun-Mei, LANG Guo-Jun, XU Wen, MAO Shan-Shan, LIN Tong   

  1. College of Forestry, South China Agricultural University, Guangzhou 510642, China
  • Received:2011-12-31 Revised:2012-03-14 Online:2012-04-20 Published:2012-04-20
  • Contact: LIN Tong E-mail: lintong@scau.edu.cn
  • About author:realein@yahoo.com.cn

Abstract: Abstract: 【Aim】 Ophiusa disjungens (Lepidoptera, Noctuidae) is one of the main insect pests that attack eucalyptus. A new nucleopolyhedrovirus (NPV) called Ophiusa disjungens NPV (OpdiNPV) was recently isolated from diseased larvae. The objective of this research was to understand OpdiNPV at the molecular level. 【Methods】 A PstⅠ genomic fragment of OpdiNPV was cloned, sequenced and analyzed. 【Results】 A homologue of Ac108 gene, assigned as Opdi108 (GenBank accession number EU 732666), was found in this fragment. The open reading frame (ORF) of Opdi108 is 225 bp in length and its encoded protein has 16%-37% amino acid identities with other known baculovirus homologous proteins. Upstream of the start codon ATG of Opdi108, contains a late promoter motif TAAG. The ORF of Opdi108 with His-tag at C-terminus was expressed as a protein in Escherichia coli with a molecular mass of 28.7 kDa. EGFP-Opdi108 fusion protein was expressed in Trichoplusia ni (Tn) cells using Bac-to-Bac system, suggesting that Opdi108 was located mainly in the cytoplasm of infected cells at 24 h post-infection (p.i.) and 72 h p.i. 【Conclusion】 This study provides a genetic basis for the development of the virus OpdiNPV as a biopesticide or an engineered pesticide to control O. disjungens and other lepidopteran insects.

Key words: Ophiusa disjungens, nucleopolyhedrovirus, Bac-to-Bac expression system, cellular localization, Opdi108