Robert Lamarche (robert at bio1.lan.mcgill.ca) wrote:
: We have introduced Jim Haseloff's 35S-GFP construct into plants. Several
: independent transformants from independent transformations display green
: fluorescence specific to the guard cells. No other green fluorescence is
: observed in the transformants; no green fluorescence is observed in any
: cell type in the controls. We have tried both FITC on the fluorescent
: microscope and confocal imaging and only find this guard cell specific
: signal. This cell specificity has surprised us. Has any one else made a
: similar observation or are we looking at some artifact? Any comments from
: people working with GFP in plants would be much appreciated.
: Candace Waddell
: Robert Lamarche, Systems Manager
: Biology Dept. McGill University
: 1205 Docteur Penfield ave., Room W2/2
: Montreal, Quebec, Canada H3A 1B1
: Phone: (514) 398-6469 Fax: (514) 398-5005
Please give use more details regarding the gene construct you used. An
observation from Beachy's lab at Scripps comes to mind where he inserted
the wt GFP gene into tobacco plants and none of them showed any
fluorescence exept one tiny green spot in one of the plants. He looked
deeper into that phenomenon and found that he had cloned some 3'
untranslated sequence together with the GFP into his expression vector.
The plasmid recovered from the green spot had a deletion of this 3'
untranslated sequence. It turned out that this sequence suppresses the
expression of GFP in his plants. It might be a tissue-specific
suppression - so please check whether you cloned the GFP ORF only or
had some UTR attached to it.
Hope that helps.
With best regards