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Extended vs. Condensed: Determination of Mitochondrial
Extended vs. Condensed: Determination of Mitochondrial

... organism for eukaryotes • A species of budding yeast • About 3-5 μm • Small genome: 1.4 x 107 basepairs ...
Plasmodesmata 2004. Surfing the Symplasm
Plasmodesmata 2004. Surfing the Symplasm

... including a group that label punctae at the cell periphery, a pattern consistent with PD localization. Macromolecules that traffic by a selective or targeted pathway might be expected to contain sequences necessary for translocation. Kimberley Gallagher (Duke University, Durham, NC) was able to show ...
Hydrophobic Interaction Chromatography (HIC
Hydrophobic Interaction Chromatography (HIC

... *Hydrophobic literally means “water fearing.” It refers to substances that do not mix well with water (i.e., Oil is hydrophobic). Note how it stays together in small beads and does not mix with water. Purpose: To separate hydrophobic proteins based on their tendency to stick to specially treated bea ...
No Slide Title
No Slide Title

... 1/3 of genome (48.5 Kb) is non essential DNA is packaged into phage particles Can only fit 40 – 53 Kb of DNA Have an in vitro packaging system Highly efficient at transforming bacteria – Can clone up to 23 Kb of DNA ...
Plant Cell and Environment
Plant Cell and Environment

... 1999). It has recently been demonstrated that several mutant green fluorescent proteins (GFPs) can be used as non-invasive intra- and extracellular pH sensors (Kneen, Farinas & Verkman 1998; Llopis et al. 1998; Miesenbock, Angelis & Rothman 1998). In particular, a pH-sensitive GFP derivative, ‘ratio ...
Acetyl chloride (A0772) - Product Information Sheet - Sigma
Acetyl chloride (A0772) - Product Information Sheet - Sigma

... corresponding thiohydantoins that incorporates acetyl chloride has been published.5 The use of acetyl chloride in the synthesis of cladribine (2-chloro-2'deoxyadenosine) from 2'-deoxyguanosine has been reported.6 Acetyl chloride can induce activation of a reporter gene which codes for a green fluore ...
CytoTracers User Manual
CytoTracers User Manual

... Molecular trafficking is a dynamic process in eukaryotic cells. The Cyto-Tracers™ provide a means of live monitoring of molecules in real-time. SBI has created a line of lentivector-based Cyto-Tracers that utilize GFP-fusion proteins to mark cellular compartments, organelles, vesicles and structures ...
BioWire_Progress_Report,_Week_One
BioWire_Progress_Report,_Week_One

... [Components will be ordered Monday] Replicate Weiss sender cell, adding GFP as a reporter Manually add aTc to the system in varying concentrations Control: Add water instead of aTc Expected Result: GFP is expressed, but not in control plates. ...
Peroxisome degradation requires catalytically active sterol
Peroxisome degradation requires catalytically active sterol

... synthesize SG and to execute pexophagy. In order to investigate the localization of the domain-deleted enzymes, green ¯uorescent protein (GFP) was fused to the N-terminus of each full-length Ugt51 protein, and the constructs were then expressed under the control of the Ugt51 promoter (strain PoGFP±U ...
Induction of fungal cell wall stress
Induction of fungal cell wall stress

... suspension was diluted to the concentration of 2 x 104 CFU/mL in the complete media (CM) [Bennett and Lasure, 1991]. One hundred μL of A. niger (RD 6.47 or JD 1.1) was added to each well after adding the plant extracts, compounds and controls. Aspergillus niger N402 was used to indicate the GFP non- ...
Chemiluminescent and Fluorescent Westerns
Chemiluminescent and Fluorescent Westerns

... resolved easily through gel electrophoresis. For example, chemiluminescence is often used to detect the induction of exogenous protein expression in transfected cell lines, to confirm specific purification of a known protein, or for verification of antibodies during production. Chemiluminescent draw ...
New degradation proteins show route to cell survival
New degradation proteins show route to cell survival

... Further studies linked the two proteins to autophagy of a certain cell constituent – the endoplasmic reticulum (ER), a network of flattened membrane enclosed sacks – in nitrogen-starved conditions. The same conditions also triggered degradation of a part of the nucleus by Atg39; this protein localiz ...
Cell-Based Applications of Living Colors® Proteins
Cell-Based Applications of Living Colors® Proteins

... screening needs. The spectral properties of fluorescent proteins make them extremely well suited for multicolor applications in fluorometry, flow cytometry, and fluorescence microscopy—fluorescent proteins have great potential in both optimizing and accelerating drug discovery and screening processe ...
Bacterial Transformation Lab
Bacterial Transformation Lab

... You will be working with a bacteria called E. coli In order to transform E. coli, you need to grow LOTS of it Today we will be placing E. coli on a petri dish and incubating it at body temperature (37◦C) to ...
Mr. David Cortens In Vivo Synthesis of ?Click? Functionalized
Mr. David Cortens In Vivo Synthesis of ?Click? Functionalized

... the copper catalysed Huisgen 1,3-dipolar cycloaddition of azides and alkynes, although other “click” reactions are possible as well (12,13). The trademark of a “click” reaction is the fact that it can easily be performed in mild, physiological conditions, without the presence of unwanted site reacti ...


... molecular weight of 27 kDa. The major spectral absorption peak is at 395 nm with a minor peak 470 nm and a single emission peak at 509 nm (Chalfie et al., 1994). Three amino acids, Ser65, Tyr66, and Gly67 associate to form the fluorophore of ...
Tonoplast and Vacuoles
Tonoplast and Vacuoles

... (PVC) before transport to the lytic vacuole C – ER-derived protein bodies filled with prolamins are autophaged by vacuoles. ...
Figure S1. Chloroplast localization and topology of TerC
Figure S1. Chloroplast localization and topology of TerC

... Figure S1. Chloroplast localization and topology of TerC-GFP fusion protein. (a) Protoplasts were isolated from terc-1TerC-GFP. Chlorophyll fluorescence was excited at 450 – 490 nm and the emission was recorded at > 515 nm (Filterset 9, Carl Zeiss, http://microscopy.zeiss.com/microscopy/en_de/servic ...
Regulation of GFP Expression
Regulation of GFP Expression

... allowing them to share these beneficial genes. This natural mechanism allows bacteria to adapt to new environments. The ability of bacteria to become resistant to antibiotics is due to their ability to accept and donate plasmids coding for antibiotic resistance from other bacterium. Scientists have ...
The symbiotic ion channel homolog DMI1 is localized in the nuclear
The symbiotic ion channel homolog DMI1 is localized in the nuclear

... an appropriate plastid targeting sequence DMI1 can be efficiently targeted to the plastids. In order to determine if GS2cTP could restore function to tDMI1::GFP, we transformed dmi1 mutants with the GS2cTP::tDMI1::GFP fusion and inoculated transgenic roots with S. meliloti. As with tDMI1::GFP, and i ...
Transient and Stable GFP Expression in Germ Cells by the vasa
Transient and Stable GFP Expression in Germ Cells by the vasa

... PGCs visualization and labeling is the first step to forward the application. They can be transiently labeled by embryonic injection of mRNA containing green fluorescence protein gene (GFP) and 3’-untranslated region (3’UTR) of a maternal germ gene such as vasa, nos1, etc. The 3’UTR sequences play a ...
蛋白質工程於生物技術 之應用與發展 Protein Engineering
蛋白質工程於生物技術 之應用與發展 Protein Engineering

... Applications in Engineering Proteins •Engineering of industrial enzymes •Re-design of substrate specificity •Folding and stability •Custom-designed proteins •Chimeric protein constructions ...
Cellular Internalization of Fluorescent Proteins via Arginine
Cellular Internalization of Fluorescent Proteins via Arginine

... compartments is limited by low biomembrane permeability because of the hydrophobic characteristics of cell membranes. Usually the delivery processes utilize passive protein channels or active transporters to overcome the membrane impediment. In this report, we demonstrate that arginine-rich intracel ...
Please read these notes before deciding what to order
Please read these notes before deciding what to order

... Please note: full preps of any of these viruses can be ordered. Inquire with me: [email protected] Please read these notes before deciding what to order: (1) “LS1L” denotes cre-dependent (2) hEF1α vectors are for long term retrograde studies. I have recently made an improvement in my packaging protocol ...
Please read these notes before deciding what to order
Please read these notes before deciding what to order

... Please note: full preps of any of these viruses can be ordered. Inquire with me: [email protected] Please read these notes before deciding what to order: (1) “LS1L” denotes cre-dependent (2) hEF1α vectors are for long term retrograde studies. I have recently made an improvement in my packaging protocol ...
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Green fluorescent protein



The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy (Renilla reniformis) has a single major excitation peak at 498 nm.In cell and molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms it has been used to make biosensors, and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, injection with a viral vector, or cell transformation. To date, the GFP gene has been introduced and expressed in many Bacteria, Yeast and other Fungi, fish (such as zebrafish), plant, fly, and mammalian cells, including human. Martin Chalfie, Osamu Shimomura, and Roger Y. Tsien were awarded the 2008 Nobel Prize in Chemistry on 10 October 2008 for their discovery and development of the green fluorescent protein.
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