Construction of gene expression systems in insect cell lines using promoters from the silkworm, Bombyx mori.
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 beta-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.
Lee
Jae Man
JM
Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki 6-10-1, Fukuoka 812-8581, Japan.
Takahashi
Masateru
M
Mon
Hiroaki
H
Mitsunobu
Hitoshi
H
Koga
Katsumi
K
Kawaguchi
Yutaka
Y
Nakajima
Yumiko
Y
Kusakabe
Takahiro
T
D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins