The green fluorescent protein (GFP) holds promise as a field-level transgene marker. One obstacle to the use of GFP is fluorescence variability observed within leaf canopies. In growth chamber and field experiments, GFP fluorescence in transgenic oilseed rape ( Brassica napus) was shown to be variable at each leaf position over time and among different leaves on the same plant. A leaf had its highest GFP fluorescence after emergence and, subsequently, its fluorescence intensity decreased. GFP fluorescence intensity was directly correlated with the concentration of soluble protein. The concentration of the genetically linked recombinant Bacillus thuringiensis (Bt) cry1Ac endotoxin protein also was examined, and GFP fluorescence was positively correlated with Bt throughout development. The results show that GFP can be used as an accurate transgene marker but that aspects of plant developmental should be taken into account when interpreting fluorescence measurements.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/s00299-003-0696-4 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Method for determining of cytotoxicity based on the release of fluorescent proteins.
BMC Mol Cell Biol
January 2025
Institute of Future Biophysics, Institutskiy per. 9, Dolgoprudny, Moscow Oblast, Moscow, Russia.
This paper describes a method for determining the cytotoxicity of chemical compounds based on the detection of fluorescent proteins-in this case, green fluorescent protein (GFP) and red fluorescent protein (RFP), which are released into the medium from dead cells. This method is similar in principle to the lactate dehydrogenase test (LDH test), but it does not require a reaction with a chromogenic substrate. This method also makes it possible to independently determine the viability of different lines when used in cocultures.
View Article and Find Full Text PDFNew insights into the regulation of cyp3a65 expression in transgenic tg(cyp3a65:GFP) zebrafish embryos.
Aquat Toxicol
January 2025
Unité écotoxicologie des substances et des milieux, Institut National de l'Environnement Industriel et des Risques (INERIS), 60550 Verneuil-en-Halatte, France. Electronic address:
Facing the need for alternative models allowing assessment of metabolic-endocrine disrupting chemicals (MDCs), especially in poorly investigated tissues such as the intestine, we recently developed a transgenic zebrafish embryo in vivo model, tg(cyp3a65:GFP), expressing the Green Fluorescent Protein (GFP) under the control of the zebrafish cyp3a65 promoter, ortholog of human cyp3a4, a gene coding for a key enzyme of intestinal xenobiotic and endobiotic metabolism. In this study, we aimed to better understand the regulation of cyp3a65 expression by zfPXR, zfAhR2, and zfGR zebrafish orthologs of well-known human xenosensors PXR and AhR, and steroid nuclear receptor GR. For this purpose, we performed zebrafish embryo tg(cyp3a65:GFP) (co)exposures to a variety of agonists (clotrimazole, TCDD, fluticasone propionate) and antagonists (econazole nitrate, CH223181, RU486), which were characterized using in vitro zebrafish reporter gene assays.
View Article and Find Full Text PDFA high-throughput, microplate reader-based method to monitor in vitro HIV latency reversal in the absence of flow cytometry.
Virology
January 2025
The Wistar Institute of Anatomy and Biology, Philadelphia, PA, USA. Electronic address:
J-Lat cells are derivatives of the Jurkat CD4 T cell line that contain a non-infectious, inducible HIV provirus with a GFP tag. While these cells have substantially advanced our understanding of HIV latency, their use by many laboratories in low and middle-income countries is restricted by limited access to flow cytometry. To overcome this barrier, we describe a modified J-Lat assay using a standard microplate reader that detects HIV-GFP expression following treatment with latency-reversing agents (LRAs).
View Article and Find Full Text PDFTrueSpot: A robust automated tool for quantifying signal puncta in fluorescent imaging.
bioRxiv
January 2025
Department of Molecular Physiology and Biophysics, School of Medicine, Vanderbilt University, Nashville, TN, 37232, USA.
Characterizing the movement of biomolecules in single cells quantitatively is essential to understanding fundamental biological mechanisms. RNA fluorescent in situ hybridization (RNA-FISH) is a technique for visualizing RNA in fixed cells using fluorescent probes. Automated processing of the resulting images is essential for large datasets.
View Article and Find Full Text PDFOxygenation and function of endocrine bioartificial pancreatic tissue constructs under flow for preclinical optimization.
J Tissue Eng
January 2025
Department of Chemical Engineering, McGill University, Montreal, QC, Canada.
Islet transplantation and more recently stem cell-derived islets were shown to successfully re-establish glycemic control in people with type 1 diabetes under immunosuppression. These results were achieved through intraportal infusion which leads to early graft losses and limits the capacity to contain and retrieve implanted cells in case of adverse events. Extra-hepatic sites and encapsulation devices have been developed to address these challenges and potentially create an immunoprotective or immune-privileged environment.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!