Abstract
The cell lines derived from the fall armyworm (FAW), Spodoptera frugiperda, have been widely used for production of recombinant proteins for applications in both basic research and applications in medicine and agriculture. Promoters from the nucleopolyhedrovirus (NPV) are commonly used in these expression systems. These promoters have some limitations, which may be overcome by using promoters of genes from S. frugiperda. However, information on these promoters is not available. We identified several highly expressed genes from the transcriptomes of S. frugiperda midgut, fat body, epidermis, ovarian cell line (Sf9), and a midgut cell line (Sf17). The activity of potential promoters of 21 highly expressed genes was evaluated in Sf9 and Sf17 cells. Two of these promoters, SfHSC70-P1780 and SfPub-P2009, showed higher activity than commonly used hr5/ie1 (baculovirus enhancer element, hr5 and immediate early gene 1, ie1) promoter. Interestingly, the activity of these two promoters increased after adding hr5 enhancer element. The hr5/SfPub-P2009 promoter performance was evaluated by expressing an exogenous P450 protein in Sf9 cells using a plasmid-based expression system. The activity of this promoter was also evaluated in the FAW by expressing green fluorescence protein using the baculovirus expression system. In both cases, the hr5/SfPub-P2009 promoter performed better than the commonly used hr5/ie1 promoter. These strong endogenous promoters will be useful for studies in S. frugiperda and other lepidopteran insects for multiple applications, including protein expression, genome editing, and transgenic insects.
Original language | English |
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Article number | 103455 |
Journal | Insect Biochemistry and Molecular Biology |
Volume | 126 |
DOIs | |
State | Published - Nov 2020 |
Bibliographical note
Funding Information:We thank Dr. Cynthia L. Goodman from USDA, ARS, BCIRL at Columbia, MO, for the gift of SfMG_0617 cells. This material is based on work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under Grant No. IIP1821936, and by industry partners, and the National Institute of Food and Agriculture, US Department of Agriculture (under HATCH Project 2353057000).
Funding Information:
We thank Dr. Cynthia L. Goodman from USDA, ARS, BCIRL at Columbia, MO, for the gift of SfMG_0617 cells. This material is based on work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under Grant No. IIP1821936 , and by industry partners, and the National Institute of Food and Agriculture, US Department of Agriculture (under HATCH Project 2353057000 ).
Publisher Copyright:
© 2020 Elsevier Ltd
Keywords
- Baculovirus and hr5 enhancer
- Protein expression
- Sf9 cells
- Ubiquitin promoter
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Insect Science