NDLI: Comparison of the MpEF1α and CaMV35 promoters for application in Marchantia polymorpha overexpression studies

Content Provider	Springer Nature Link
Author	Althoff, Felix Kopischke, Sarah Zobell, Oliver Ide, Kentaro Ishizaki, Kimitsune Kohchi, Takayuki Zachgo, Sabine
Copyright Year	2013
Abstract	Constitutive promoters are essential tools for analyses of gene functions by transgenic approaches. For overexpression and silencing studies of genes, a ubiquitous and strong expression of genes under investigation as well as selection markers is preferred. For future applications in the emerging basal plant model system Marchantia polymorpha, a liverwort, activities of the viral 35S cauliflower mosaic virus promoter and the endogenous elongation factor 1α (MpEF1α) promoter were analyzed. Expression of the reporter gene β-glucuronidase (GUS), driven by the CaMV35 and MpEF1α promoters, was compared throughout plant development. Significant differences were observed between the two promoter activities. The CaMV35 promoter yields a weak reporter gene expression in the meristematic zones but drives a strong expression in the thallus. The MpEF1α promoter causes a strong meristematic GUS expression and is more active in female sexual tissues. Overall, the MpEF1α promoter seems to be the better option for obtaining a strong and ubiquitous transgene expression. Furthermore, a whole mount in situ hybridization protocol for Marchantia was established. Analysis of MpEF1α mRNA transcript in intact, whole tissues showed an expression pattern that is overall similar to the pattern of the GUS reporter gene expression driven by the MpEF1α promoter, including strong expression in meristematic zones. The whole mount technique reported here can be used to determine the mRNA expression in intact gemmae and archegonia, and has the potential to be applied for screening large numbers of transgenic plants, for instance to identify knock-down mutants.
Starting Page	235
Ending Page	244
Page Count	10
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	23
Issue Number	2
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2013-09-15
Publisher Place	Cham
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	CaMV35 promoter EF1α promoter Marchantia polymorpha β-glucuronidase staining In situ whole mount hybridization Molecular Medicine Plant Genetics & Genomics Animal Genetics and Genomics Plant Sciences Human Genetics
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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