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Journal of Biochemistry Advance Access originally published online on November 4, 2008
Journal of Biochemistry 2009 145(1):37-42; doi:10.1093/jb/mvn147
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© The Authors 2008. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved

Production and Characterization of the Recombinant Human µ-Opioid Receptor from Transgenic Silkworms

Mayuko Tateno1, Masaru Toyooka1, Yohko Shikano1, Shigeki Takeda1,*, Nobuo Kuwabara2, Hideki Sezutsu3 and Toshiki Tamura3

1Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan; 2Gunma Sericultural Technology Center, 1-1-1 Ohte-machi, Maebashi, Gunma 371-8570, Japan; and 3Transgenic Silkworm Research Center, National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan

*To whom correspondence should be addressed. Tel/Fax: +81-277-30-1434, E-mail: stakeda{at}chem-bio.gunma-u.ac.jp

Received September 24, 2008; Accepted October 14, 2008


  Abstract

The production of useful quantities of G protein-coupled receptors is a major problem not only for screening of various drug compounds but also in performing structural biology studies. To solve this problem, we investigated the possibility of using transgenic silkworms for the production of these receptors. Using the human µ-opioid receptor gene, we constructed three transgenic silkworm strains that produced µ-opioid receptors. The silkworms expressed significant amounts of the receptor in the fat body and silk gland. The product was evaluated using a saturation ligand-binding assay. The expressed receptor exhibited ligand affinity similar to that of an authentic sample, and the yield from the transgenic silkworm was comparable to that obtained using an Sf9-baculovirus expression system. As the mass rearing of transgenic silkworms has already been established, the silkworms can be adapted for production of large quantities of receptors.

Key Words: G protein-coupled receptor, opioid receptor, piggyBac transposon elements, transgenic silkworm, UAS-Gal4 system

Abbreviations: A3 promoter, silkworm cytoplasmic actin gene promoter; GPCRs, G protein-coupled receptors; UAS, upstream activating sequence for Gal4; IVR, inverse repeat sequence


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