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

Scott F Cummins; A Thongkukiatkul; P J Hanna

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

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

Scott F Cummins, A Thongkukiatkul and P J Hanna

Journal of Shellfish Research, Vol.20(2), pp.705-710

2001

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Abstract

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 of polyclonal 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 of neural 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.

Details

Title: Location of egg-laying hormone in reproductive structures and neurons of Haliotis rubra (Leach) using antibodies raised against recombinant fusion proteins
Authors: Scott F Cummins (Author) - Deakin University
A Thongkukiatkul (Author) - Burapha University, Thailand
P J Hanna (Author) - Deakin University
Publication details: Journal of Shellfish Research, Vol.20(2), pp.705-710
Publisher: National Shellfisheries Association, Inc.
Date published: 2001
ISSN: 0730-8000
Organisation Unit: School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 99451194102621
Output Type: Journal article

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