Premium
Green fluorescent protein as a reporter system in the transformation of barley cultivars
Author(s) -
Ahlandsberg Staffan,
Sathish P.,
Sun Chuanxin,
Jansson Christer
Publication year - 1999
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1034/j.1399-3054.1999.100207.x
Subject(s) - transformation (genetics) , cultivar , fluorescence , green fluorescent protein , reporter gene , fluorescent protein , chemistry , botany , biology , biochemistry , gene , gene expression , optics , physics
A cereal transformation vector, pN1473, containing the strong constitutive rice actin promoter Act‐1 , a multiple cloning site, and the nos terminator, was constructed. Fusion of a plant‐optimized gfp gene to Act‐1 in pN1473 resulted in the vector pN1473GFP. To assess the suitability of pN1473, and GFP as a reporter system in barley transformation, two barley cultivars (Baronesse and Golden Promise) were transformed by microprojectile bombardment. Transient gfp expression in transformed embryogenic callus material was detectable by fluorescence microscopy less than 12 h after transformation. The presence of the gfp gene in callus and regenerated plantlets was confirmed by PCR amplification and DNA gel‐blot analysis.