Construction of gene expression systems in insect cell lines using promoters from the silkworm, Bombyx mori

Jae Man Lee, Masateru Takahashi, Hiroaki Mon, Hitoshi Mitsunobu, Katsumi Koga, Yutaka Kawaguchi, Yumiko Nakajima, Takahiro Kusakabe

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The promoter regions of the Bombyx mori HSC70-4 and B. mori TCTP genes characterized previously were used for the construction of a series of constitutive gene expression systems active in cultured cells. The relative abilities of these promoters were evaluated by comparing those of a silkworm actin A3 (BmActin3) promoter, which is used widely as the first choice. A series of constitutive expression systems constructed were assayed for the transcription efficiency by connecting four reporter cDNAs, firefly luciferase, 3GFP, Ds-Red, and β-galactosidase gene using the Gateway LR reaction. The insertion of an intron enhancer into the site between the TCTP promoter and gene increased the transcription of the BmTCTP promoter by 10-fold. The insertion of the IE-1 gene and HR3 enhancer to the all three promoters were found to increase the transcription up to 560 times.

Original languageEnglish
Pages (from-to)9-17
Number of pages9
JournalJournal of Biotechnology
Volume133
Issue number1
DOIs
Publication statusPublished - Jan 1 2008

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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