期刊
MARINE GENOMICS
卷 42, 期 -, 页码 41-48出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.margen.2018.06.003
关键词
Diatoms; Phaeodactylum tricornutum; V-ATPase C promoter; High-level expression; Constitutive expression
资金
- Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan [15K14804]
- Grants-in-Aid for Scientific Research [15K14804] Funding Source: KAKEN
The marine diatom Phaeodactylum tricornutum is attractive for basic and applied diatom research. We isolated putative endogenous gene promoters derived from genes that are highly expressed in P. tricornutum: the fucoxanthin chlorophyll a/c-binding protein (FCP) C gene, the vacuolar ATP synthase 16-kDa proteolipid subunit (V-ATPase C) gene, the clumping factor A gene and the solute carrier family 34 member 2 gene. Five putative promoter regions were isolated, linked to an antibiotic resistance gene (Sh ble) and transformed into P. tricomutum. Using quantitative RT-PCR, the promoter activities in the transformants were analyzed and compared to that of the diatom endogenous gene promoter, the FCP A gene promoter which has been used for the transformation of P. tricornutum. Among the five isolated potential promoters, the activity of the V-ATPase C gene promoter was approximately 2.73 times higher than that of the FCP A gene promoter. The V-ATPase C gene promoter drove the expression of Sh ble mRNA transcripts under both light and dark conditions at the stationary phase. These results suggest that the V-ATPase C gene promoter is a novel tool for the genetic engineering of P. tricornutum.
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