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

Citation
T. Ponappa et al., Transient expression and stable transformation of soybean using the jellyfish green fluorescent protein, PL CELL REP, 19(1), 1999, pp. 6-12
Citations number
25
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT CELL REPORTS
ISSN journal
0721-7714 → ACNP
Volume
19
Issue
1
Year of publication
1999
Pages
6 - 12
Database
ISI
SICI code
0721-7714(199911)19:1<6:TEASTO>2.0.ZU;2-L
Abstract
Embryogenic soybean [Glycine max (L,) Merrill.] suspension cultures were bo mbarded with bye different gene constructions encoding the jellyfish (Aequo rea victoria) green fluorescent protein (GFP). These constructions had alte red codon usage compared to the native GFP gene and mutations that increase d the solubility of the protein and/or altered the native chromophore. All of the constructions produced green fluorescence in soybean cultures upon b lue light excitation, although a soluble modified red-shifted GFP (smRS-GFP ) was the easiest to detect based on the brightness and number of foci prod uced. Expression of smRS-GFP was visible as early as 1.5 h after bombardmen t, with peak expression at approximately 6.5 h, Large numbers of smRS-GFP-e xpressing areas were visible for 48 h postbombardment and declined rapidly thereafter. Stably transformed cultures and plants exhibited variation in t he intensity and location of GFP expression. PCR and Southern hybridization analyses confirmed the presence of introduced GFP genes in stably transfor med cultures.