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

doi:10.1016/j.jbiotec.2007.08.033

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^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

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 language	English (US)
Pages (from-to)	9-17
Number of pages	9
Journal	Journal of Biotechnology
Volume	133
Issue number	1
DOIs	https://doi.org/10.1016/j.jbiotec.2007.08.033
State	Published - Jan 1 2008
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN), and by a grants-in-aid no. 17380037 from the Ministry of Education, Science and Culture of Japan.

Keywords

Constitutive expression system
Enhancer
Gateway LR reaction
Reporter cDNAs
Transcription efficiency

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1016/j.jbiotec.2007.08.033

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

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.",

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note = "Funding Information: This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN), and by a grants-in-aid no. 17380037 from the Ministry of Education, Science and Culture of Japan.",

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AU - Koga, Katsumi

AU - Kawaguchi, Yutaka

AU - Nakajima, Yumiko

AU - Kusakabe, Takahiro

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KW - Transcription efficiency

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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