Characterization and Sequence Determination of Locust Brain cDNA Clones Selected by Antisera Raised Against Vertebrate Peptide Hormones
By means of immunocytochemical studies, performed with antisera directed against known (vertebrate) peptide hormones, numerous immunopositive substances have been detected within insect nervous tissue and gut endocrine cells. The isolation and characterization of these immunoreactive substances, however, may become a very laborious task. Therefore, an alternative and complementary approach was tried. A locust (Locusta migratoria) brain cDNA expression library was constructed. This recombinant DNA approach enabled the use of antisera as probes for the selection of specific nucleic acid sequences. In this study bovine growth hormone- (bGH-) and melanotropin- (MSH-), as well as hypertrehalosemic factor- (HTF- from Carausius morosus) and caudo-dorsal cell hormone- (CDCH- from Lymnea stagnaliss) immunopositive plaques have been isolated, purified and amplified. These phage clones were then further analyzed. The sequence of cDNA inserts has been determined.
This immunological screening approach, combined with hybridization analysis of the isolated clones, can lead to the characterization of nucleic acid sequences coding for insect tissue immuno-reactive substances.
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