Expression of neuroparsin cDNA in insect cells using baculovirus vectors.
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.
Girardie
J
J
Laboratoire de Neuroendocrinologie, URA CNRS 1138, Université Bordeaux I, Talence, France. j.girardie@invertebre.u-bordeaux.fr
Chaabihi
H
H
Fournier
B
B
Lagueux
M
M
Girardie
A
A
D000818:Animals; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics*; D004231:Diuresis / Q000187:drug effects; D015870:Gene Expression; D017344:Genes, Insect; D005822:Genetic Vectors; D006110:Grasshoppers / Q000235:genetics; D007301:Insect Hormones / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D012007:Rectum / Q000187:drug effects; D018411:Spodoptera / Q000235:genetics
D018076:DNA, Complementary; D007301:Insect Hormones; D011994:Recombinant Proteins; C058919:neuroparsin A