Synthetic pheromone biosynthesis activating neuropeptide gene expressed in a baculovirus expression system

Insect Biochem Mol Biol. 1995 May;25(5):583-9. doi: 10.1016/0965-1748(94)00097-2.

Abstract

A synthetic gene of the pheromone biosynthesis activating neuropeptide (PBAN) of corn earworm Helicoverpa zea, with and without a signal sequence of the cuticle protein of Drosophila melanogaster, was cloned behind the polyhedrin promoter of AcMNPV. Two recombinant baculoviruses were constructed and used to infect a number of insect cell lines including Sf9 and 5B1-4. High pheromonotropic activity was consistently obtained from 5B1-4 cell culture that was infected with the recombinant baculovirus vINV-4 containing the signal sequence. The PBAN gene-product was isolated by HPLC and analyzed by electrospray ionization mass spectrometry. Low levels of biological activity obtained from Sf9 cells infected with the recombinant virus vPBAN may be due to lack of proper amidation at the C-terminus of the expressed peptide or rapid proteolytic degradation of the product.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • DNA
  • Drosophila melanogaster / genetics
  • Molecular Sequence Data
  • Neuropeptides / genetics*
  • Neuropeptides / metabolism
  • Nucleopolyhedroviruses / genetics*
  • Protein Sorting Signals / genetics
  • Recombinant Fusion Proteins / genetics
  • Sex Attractants / genetics*
  • Sex Attractants / metabolism
  • Spodoptera

Substances

  • Neuropeptides
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Sex Attractants
  • pheromone biosynthesis activating neuropeptide, Helicoverpa zea
  • DNA