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

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

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)375-379
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume385
Issue number3
DOIs
Publication statusPublished - Jul 31 2009

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Bombyx
G-Protein-Coupled Receptors
Nucleopolyhedrovirus
DNA
Fusion reactions
Baculoviridae
GTP-Binding Protein alpha Subunits
Guanine Nucleotides
Centrifugation
Sf9 Cells
Carrier Proteins
Proteins
Fat Body
Fats
Larva
nociceptin receptor
Injections

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system. / Kajikawa, Mizuho; Tabata, Kaori; Wakimoto, Yoshitaro; Toyooka, Masaru; Motohashi, Tomoko; Shimojima, Tsukasa; Takeda, Shigeki; Park, Enoch Y.; Maenaka, Katsumi.

In: Biochemical and Biophysical Research Communications, Vol. 385, No. 3, 31.07.2009, p. 375-379.

Research output: Contribution to journalArticle

Kajikawa, M, Tabata, K, Wakimoto, Y, Toyooka, M, Motohashi, T, Shimojima, T, Takeda, S, Park, EY & Maenaka, K 2009, 'Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system', Biochemical and Biophysical Research Communications, vol. 385, no. 3, pp. 375-379. https://doi.org/10.1016/j.bbrc.2009.05.063
Kajikawa, Mizuho ; Tabata, Kaori ; Wakimoto, Yoshitaro ; Toyooka, Masaru ; Motohashi, Tomoko ; Shimojima, Tsukasa ; Takeda, Shigeki ; Park, Enoch Y. ; Maenaka, Katsumi. / Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system. In: Biochemical and Biophysical Research Communications. 2009 ; Vol. 385, No. 3. pp. 375-379.
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