Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system

Mizuho Kajikawa, Kaori Sasaki, Yoshitaro Wakimoto, Masaru Toyooka, Tomoko Motohashi, Tsukasa Shimojima, Shigeki Takeda, Enoch Y. Park, Katsumi Maenaka

研究成果: Contribution to journalArticle査読

11 被引用数 (Scopus)

抄録

Guanine nucleotide-binding protein (G protein) coupled receptors (GPCRs) are frequently expressed by a baculovirus expression vector system (BEVS). We recently established a novel BEVS using the bacmid system of Bombyx mori nucleopolyhedrovirus (BmNPV), which is directly applicable for protein expression in silkworms. Here, we report the first example of GPCR expression in silkworms by the simple injection of BmNPV bacmid DNA. Human nociceptin receptor, an inhibitory GPCR, and its fusion protein with inhibitory G protein alpha subunit (Giα) were both successfully expressed in the fat bodies of silkworm larvae as well as in the BmNPV viral fraction. Its yield was much higher than that from Sf9 cells. The microsomal fractions including the nociceptin receptor fusion, which are easily prepared by only centrifugation steps, exhibited [35S]GTPγS-binding activity upon specific stimulation by nociceptin. Therefore, this rapid method is easy-to-use and has a high expression level, and thus will be an important tool for human GPCR production.

本文言語英語
ページ(範囲)375-379
ページ数5
ジャーナルBiochemical and Biophysical Research Communications
385
3
DOI
出版ステータス出版済み - 7 31 2009

All Science Journal Classification (ASJC) codes

  • 生物理学
  • 生化学
  • 分子生物学
  • 細胞生物学

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