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.
Bibliographical noteFunding 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.
- Constitutive expression system
- Gateway LR reaction
- Reporter cDNAs
- Transcription efficiency
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology