Expression of neuroparsin cDNA in insect cells using Baculovirus vectors

The cDNA encoding neuroparsin A, a polytropic neurohormone of the locust, L ocusta migratoria, was inserted into the genome of Autographa californica n uclear polyhedrosis virus such that transcription was under control of the p10 promoter. A polypeptide having the same charge and the same apparent mo lecular weight as the authentic neuroparsin A and that was reactive against neuroparsin immune serum was produced in recombinant virus-infected lepido pteran cell lines but not in control virus-infected cells. The baculovirus- expressed polypeptide was purified by two steps of liquid chromatography (a nion exchange and reversed phase) which were previously used to purify the natural neuroparsin. The purified baculovirus-expressed polypeptide enhance d fluid reabsorption of everted rectum preparations, as did the natural neu roparsin, Thus, this gene expression system produced a polypeptide identica l to authentic neuroparsin, (C) 1997 Wiley-Liss, Inc.