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Stable Genetic Transformation of Eschscholzia Californica Expressing Synthetic Green Fluorescent Proteins
Author(s) -
Lee Jintae,
Pedersen Henrik
Publication year - 2001
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp010003v
Subject(s) - agrobacterium tumefaciens , poppy , transformation (genetics) , green fluorescent protein , biology , cauliflower mosaic virus , fluorescence , agarose , fluorescent protein , microbiology and biotechnology , reporter gene , botany , gene , genetics , genetically modified crops , gene expression , transgene , physics , quantum mechanics
Abstract An efficient protocol is described for the stable genetic transformation of Eschscholzia californica (California poppy) using Agrobacterium tumefaciens as a vector. We have employed the disarmed A. tumefaciens LBA4404 encoding a synthetic green fluorescent protein reporter gene that is further controlled by an enhanced cauliflower mosaic virus 35 S promoter. Stably transformed E. californica cells appear 3 weeks after initial cocultivation of A. tumefaciens with poppy leaves, stems, or roots. Transformed poppy calli were visualized by exposure to long‐wave UV or blue light and analyzed in detail by fluorescent microscopy and laser‐scanning microscopy. Moreover, green fluorescent calli have been maintained through continual subculture and grow well either on Gamborg's B5 agarose or liquid medium.