Expression of the green fluorescent protein (gfp) gene, under regulatory control of either the constitutive 35S promoter or the developmentally-regulated lectin promoter was monitored and quantified using a newly-developed automated tracking system. The automated system consisted of a computer-controlled two-dimensional robotics table and a programmable image acquisition system, which was used to semi-continuously monitor gfp gene expression during development of transgenic soybean [Glycine max (L.) Merrill] somatic embryos. Quantitative analysis of GFP expression showed that, during somatic embryo proliferation and early development, expression of lectin-GFP was not detected. During late embryo development, expression of lectin-GFP gradually increased until the levels were similar to those of 35S-GFP. The use of an automated image collection system and image analysis facilitated the frequent monitoring and quantification of gfp gene expression on a large number of samples over an extended period of time.
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We would like to acknowledge Jim Haseloff for the gift of the mgfp5-ER gene and Lila Vodkin for kindly providing the pGLe10 plasmid containing the lectin promoter. Salaries and research support were provided by State and Federal funds appropriated to OSU/OARDC, by the United Soybean Board, and from a scholarship from the Mexican government (CONACYT) to MTBN. Mention of trademark or proprietary products does not constitute a guarantee or warranty of the product by OSU/OARDC and also does not imply approval to the exclusion of other products that may also be suitable. H&CS article #HCS 06-02.
Communicated by M. C. Jordan
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Buenrostro-Nava, M.T., Ling, P.P. & Finer, J.J. Comparative analysis of 35S and lectin promoters in transgenic soybean tissue using an automated image acquisition system and image analysis. Plant Cell Rep 25, 920–926 (2006) doi:10.1007/s00299-006-0142-5
- Green fluorescent protein
- Image analysis
- Promoter analysis