Transient expression and stable transformation of soybean using the jellyfish green fluorescent protein

Embryogenic soybean [Glycine max (L.) Merrill.] suspension cultures were bombarded with five different gene constructions encoding the jellyfish (Aequorea victoria) green fluorescent protein (GFP). These constructions had altered codon usage compared to the native GFP gene and mutations that increas...

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Bibliographic Details
Published in:Plant cell reports 1999-11, Vol.19 (1), p.6-12
Main Authors: Ponappa, T, Brzozowski, A.E, Finer, J.J
Format: Article
Language:English
Subjects:
Aequorea victoria
cell suspension culture
developmental stages
fluorescent tracers
gene expression
gene transfer
genes
genetic markers
genetic transformation
Glycine max
green fluorescent protein
plant morphology
proteins
Scyphozoa
Soybeans
timing
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Summary:Embryogenic soybean [Glycine max (L.) Merrill.] suspension cultures were bombarded with five different gene constructions encoding the jellyfish (Aequorea victoria) green fluorescent protein (GFP). These constructions had altered codon usage compared to the native GFP gene and mutations that increased the solubility of the protein and/or altered the native chromophore. All of the constructions produced green fluorescence in soybean cultures upon blue light excitation, although a soluble modified red-shifted GFP (smRS-GFP) was the easiest to detect based on the brightness and number of foci produced. Expression of smRS-GFP was visible as early as 1.5 h after bombardment, with peak expression at approximately 6.5 h. Large numbers of smRS-GFP-expressing areas were visible for 48 h postbombardment and declined rapidly thereafter. Stably transformed cultures and plants exhibited variation in the intensity and location of GFP expression. PCR and Southern hybridization analyses confirmed the presence of introduced GFP genes in stably transformed cultures.
ISSN:0721-7714
1432-203X
DOI:10.1007/s002990050702