Location of egg-laying hormone in reproductive structures and neurons of Haliotis rubra (Leach) using antibodies raised against recombinant fusion proteins

摘要

Recombinant abalone egg-laying hormone was produced using a bacterial expression vector. This required directional cloning of a 108-bp abalone egg-laying hormone (aELH) gene sequence, using PCR of genomic DNA with primers incorporating restriction enzyme sites, into a pGEX-2T vector. Following transformation of the recombinant vector into Escherichia coli, a GST:aELH fusion protein was produced in large quantities and then purified. This fusion peptide was used to immunize mice for the production ofpolyclonal and monoclonal antibodies, which were subsequently tested for specificity using ELISA's and Western blots. Antisera and two IgM monoclonal antibodies Were shown to react with aELH. These antibodies were used in immunocytochemistry studies ofneural and gonad tissues of sexually mature female abalone. The aELH was found to be located in neurosecretory cells of cerebral and pleuro-pedal ganglia, statocysts, and trabeculae in female gonads.