Expression of neuroparsin cDNA in insect cells using baculovirus vectors

Arch Insect Biochem Physiol. 1997;36(1):11-23. doi: 10.1002/(SICI)1520-6327(1997)36:1<11::AID-ARCH2>3.0.CO;2-Z.

Abstract

The cDNA encoding neuroparsin A, a polytropic neurohormone of the locust, Locusta migratoria, was inserted into the genome of Autographa californica nuclear polyhedrosis virus such that transcription was under control of the p10 promoter. A polypeptide having the same charge and the same apparent molecular weight as the authentic neuroparsin A and that was reactive against neuroparsin immune serum was produced in recombinant virus-infected lepidopteran cell lines but not in control virus-infected cells. The baculovirus-expressed polypeptide was purified by two steps of liquid chromatography (anion exchange and reversed phase) which were previously used to purify the natural neuroparsin. The purified baculovirus-expressed polypeptide enhanced fluid reabsorption of everted rectum preparations, as did the natural neuroparsin. Thus, this gene expression system produced a polypeptide identical to authentic neuroparsin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • DNA, Complementary / genetics*
  • Diuresis / drug effects
  • Gene Expression
  • Genes, Insect
  • Genetic Vectors
  • Grasshoppers / genetics
  • Insect Hormones / genetics*
  • Insect Hormones / isolation & purification
  • Insect Hormones / pharmacology
  • Nucleopolyhedroviruses / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology
  • Rectum / drug effects
  • Spodoptera / genetics

Substances

  • DNA, Complementary
  • Insect Hormones
  • Recombinant Proteins
  • neuroparsin A