Tateno Mayuko, Toyooka Masaru, Shikano Yohko, Takeda Shigeki, Kuwabara Nobuo, Sezutsu Hideki, Tamura Toshiki
Department of Chemistry and Chemical Biology, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan.
J Biochem. 2009 Jan;145(1):37-42. doi: 10.1093/jb/mvn147. Epub 2008 Nov 4.
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 mu-opioid receptor gene, we constructed three transgenic silkworm strains that produced mu-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.
生产大量有用的G蛋白偶联受体不仅是筛选各种药物化合物的主要问题,也是进行结构生物学研究的主要问题。为了解决这个问题,我们研究了利用转基因蚕生产这些受体的可能性。我们使用人类μ-阿片受体基因构建了三种能产生μ-阿片受体的转基因蚕品系。这些蚕在脂肪体和丝腺中大量表达该受体。使用饱和配体结合试验对产物进行了评估。所表达的受体表现出与真实样品相似的配体亲和力,并且转基因蚕的产量与使用Sf9-杆状病毒表达系统获得的产量相当。由于已经建立了转基因蚕的大规模饲养方法,这些蚕可用于大量生产受体。