Measurement of green fluorescent protein concentration in single cells by image analysis.
The gene encoding the green fluorescent protein (GFP) has been widely used in studies of gene expression. The GFP can be detected nondestructively in living cells or tissues by the green fluorescence of the protein under blue light. Solutions of enhanced GFP (EGFP) of known concentration were filled in glass capillaries and used to calibrate a method for quantitative determination of EGFP or GFP-S65T in plant cells. Images captured by a digital camera were analyzed to determine the linear range for measurement of EGFP expression. The value of the method was illustrated by analysis of the relative levels of GFP expression under control of different promoters in aleurone cells of barley.
Furtado
Agnelo
A
Cooperative Research Centre for Molecular Plant Breeding, Centre for Plant Conservation, Genetics, Southern Cross University, Military Road, Lismore, NSW 2480, Australia.
Henry
Robert
R
D000040:Abscisic Acid / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics; D005898:Glass; D049452:Green Fluorescent Proteins; D001467:Hordeum / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics*; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D012639:Seeds / Q000166:cytology / Q000235:genetics; D013997:Time Factors
D008164:Luminescent Proteins; D010940:Plant Proteins; D049452:Green Fluorescent Proteins; D000040:Abscisic Acid