Molecular Biology and Mutation of Green Fluorescent Protein

Zacharias, David A.; Tsien, Roger Y.

doi:10.1002/0471739499.ch5

Cited by 73 publications

(78 citation statements)

References 132 publications

(246 reference statements)

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“…In addition to improving maturation rates at 37°C, redesigning the nucleic acid sequence to coincide with codon preferences of the host organism has also improved the utility of GFP derivatives (Yang et al 1996;Zacharias and Tsien 2006). For example, more than 190 silent mutations were introduced into the coding sequence to optimize expression in human tissues (Yang et al 1996).…”

Section: Mutations That Improve Use Of Fps For Mammalian Systemsmentioning

confidence: 99%

Fluorescent Protein Tracking and Detection: Fluorescent Protein Structure and Color Variants

Rizzo¹,

Davidson²,

Piston³

2009

Cold Spring Harb Protoc

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INTRODUCTIONThe rapidly growing arsenal of genetically encoded fluorescent proteins (FPs) obtained from sea creatures has launched and fueled a revolution in live cell imaging. The diverse array of applications benefiting from FPs ranges from markers targeted at organelles and protein fusions designed to monitor intracellular dynamics to reporters of transcriptional regulation and in vivo probes for whole-body imaging and detection of cancer. FPs have enabled the creation of highly specific biosensors to monitor a wide range of intracellular phenomena, including pH and metal-ion concentration, protein kinase activity, apoptosis, membrane voltage, cyclic nucleotide signaling, and tracing neuronal pathways. The purpose of this article is to provide a description of the wide variety of FPs that are currently available.

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Section: Mutations That Improve Use Of Fps For Mammalian Systemsmentioning

confidence: 99%

Fluorescent Protein Tracking and Detection: Fluorescent Protein Structure and Color Variants

Rizzo¹,

Davidson²,

Piston³

2009

Cold Spring Harb Protoc

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“…The development of fluorescent proteins and luciferase enzymes as efficient in vivo imaging reporters began with biochemical characterization studies in the 1970s. The cloning of GFP allowed investigators subsequently to mutate it, which has generated multiple forms of GFP, enhanced for imaging in vivo (149)(150)(151). Thus, a monomeric GFP with increased expression, enhanced fluorescence in mammalian cells, and a single excitation maximum (l ex ) centered at a wavelength of 490 nm and an emission maximum (l em ) at 515 nm has been widely used.…”

Section: Imaging Metastases and Metastatic Potential With Optical Tecmentioning

confidence: 99%

Molecular Imaging of Metastatic Potential

Winnard¹,

Pathak²,

Dhara

et al. 2008

J Nucl Med

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If molecular imaging is to prove clinically useful it will have to surpass current, primarily anatomic techniques in terms of sensitivity and the ability to detect minimal changes in tissue. One of the most important tests for molecular imaging is to determine whether it can image the metastatic potential of tumors. Like all predictive endeavors, the imaging of such ''potential'' is a daunting task, but one that only molecular imaging-rather than standard, anatomic techniques-is likely to solve. Although difficult, imaging of metastatic potential is also arguably the most important task for molecular imaging of cancer because it is generally the dissemination of malignant tissue, not its prolonged residence in an inopportune site, which kills the patient. Below are examples of uses of molecular imaging of metastases as well as of metastatic potential, the former being a far more developed area of clinical inquiry.

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“…GFP is small, non-toxic, and easily visualized with most fluorescent microscopes. Mutation of the original sequence has provided derivatives with enhanced properties, such as increased thermostability, increased expression, and optimized excitation peaks (12). For co-labeling studies requiring the use of multiple fluorescent proteins, yellow fluorescent proteins (YFPs) and red fluorescent proteins (RFPs) have both proven useful.…”

Section: Fluorescent Proteinsmentioning

confidence: 99%