Construction of gateway-based destination vectors for detecting subcellular localization of proteins in the silkworm, Bombyx mori

Hitoshi Mitsunobu, Kosuke Sakashita, Hiroaki Mon, Hiroko Yoshida, Man Lee, Yutaka Kawaguchi, Katsumi Koga, Takahiro Kusakabe

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We have constructed 3 kinds of Gateway-based vectors applicable for the subcellular localization analysis of proteins in cultured Bombyx mori cells. Two were designed to visualize the location of an N- or C-terminal fusion protein with EGFP, expressed under the control of the baculoviral IE2 promoter. Another could express an N-terminal fusion protein with DsRed, similarly controlled by IE2. In order to test the applicability, the vectors were individually modified to have a cDNA encoding B. mori Actin1, histone H3 or TMS-1 (ubiquitous family of membrane protein), as well as firefly luciferase. As expected, these proteins showed cytosolic, nuclear, plasma membranous and putative peroxisomal distribution, respectively.

Original languageEnglish
Pages (from-to)141-145
Number of pages5
JournalJournal of Insect Biotechnology and Sericology
Volume75
Issue number3
Publication statusPublished - Oct 2006

Fingerprint

Bombyx
Bombyx mori
silkworms
Proteins
Fusion reactions
Firefly Luciferases
proteins
Histones
Membrane Proteins
luciferase
Complementary DNA
histones
membrane proteins
Plasmas
Silkworm
Destination
Localization
Protein
Membranes
testing

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Insect Science
  • Business, Management and Accounting(all)
  • Industrial and Manufacturing Engineering
  • Polymers and Plastics

Cite this

Construction of gateway-based destination vectors for detecting subcellular localization of proteins in the silkworm, Bombyx mori. / Mitsunobu, Hitoshi; Sakashita, Kosuke; Mon, Hiroaki; Yoshida, Hiroko; Lee, Man; Kawaguchi, Yutaka; Koga, Katsumi; Kusakabe, Takahiro.

In: Journal of Insect Biotechnology and Sericology, Vol. 75, No. 3, 10.2006, p. 141-145.

Research output: Contribution to journalArticle

Mitsunobu, Hitoshi ; Sakashita, Kosuke ; Mon, Hiroaki ; Yoshida, Hiroko ; Lee, Man ; Kawaguchi, Yutaka ; Koga, Katsumi ; Kusakabe, Takahiro. / Construction of gateway-based destination vectors for detecting subcellular localization of proteins in the silkworm, Bombyx mori. In: Journal of Insect Biotechnology and Sericology. 2006 ; Vol. 75, No. 3. pp. 141-145.
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