Molecular Characterization of the Actin Gene and 5′ Flanking Sequence from Brown Macroalga Saccharina japonica (Laminariales, Phaeophyta)
Loading...
Date
2024
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
MDPI
Abstract
The brown macroalga Saccharina japonica (Laminariales, Phaeophyta) is the most productive
cultured seaweed in the world. In order to improve the biosafety of transgenic kelp, it is necessary
to develop endogenous constitutive promoters, replacing those of virus origin. In this study, the
housekeeping actin gene from S. japonica (SjACT) was found to contain three exons and two introns,
representing a unique actin gene structure pattern in brown algae. Additionally, the 5′ upstream
region was obtained using genome walking, and fused to the reporter gene lacZ or EGFP to construct
promoter-detective vectors. Using an established genetic transformation system, kelps in different
life-cycle stages were transformed. The detection results showed that, in the diploid sporophyte
stage, the transient expression from the lacZ gene could be observed in the frond, stipe, or holdfast
of kelps, indicating a manner of being non-tissue-specific. And, in the haploid gametophyte stage
of S. japonica, the fluorescence of the expressed EGFP were detected in vivo in gametophyte cells of
both genders. These results indicate that the promoter of the SjACT gene (pSjACT) functions in a
constitutive manner and is expected to be a key endogenous element in the genetic manipulation
of kelps.
Description
Keywords
actin, Biolistic, GFP, lacZ, constitutive promoter, Saccharina japonica, transient expression