scholarly journals A FRET-Facilitated Photoswitching Using an Orange Fluorescent Protein with the Fast Photoconversion Kinetics

2012 ◽  
Vol 134 (36) ◽  
pp. 14789-14799 ◽  
Author(s):  
Oksana M. Subach ◽  
David Entenberg ◽  
John S. Condeelis ◽  
Vladislav V. Verkhusha
Keyword(s):  
Fluorescent Protein ◽  
Orange Fluorescent Protein

scholarly journals An orange fluorescent protein tagging system for real-time pollen tracking

2013 ◽  
Vol 6 (1) ◽  
Author(s):  
J Hollis Rice ◽  
Reginald J Millwood ◽  
Richard E Mundell ◽  
Orlando D Chambers ◽  
Laura L Abercrombie ◽  
...  
Keyword(s):  
Real Time ◽  
Fluorescent Protein ◽  
Orange Fluorescent Protein ◽  
Protein Tagging ◽  
Tagging System

Phytofluors: Phytochrome-Based Orange Fluorescent Protein Probes

1999 ◽  
Vol 5 (S2) ◽  
pp. 1050-1051
Author(s):  
J. Clark Lagarias ◽  
Beronda L. Montgomery ◽  
John T. Murphy ◽  
Shu-Hsing Wu
Keyword(s):  
Room Temperature ◽  
Fluorescent Protein ◽  
Protein Complexes ◽  
Quantum Yields ◽  
Color Intensity ◽  
Photosynthetic Organisms ◽  
Orange Fluorescent Protein ◽  
Pigment Protein Complexes ◽  
Covalently Linked ◽  
Light Color

Plants sense the light environment using pigment-protein complexes that discriminate light color, intensity, duration and direction. The most well-studied of these photoreceptors are the phytochromes, a family of soluble biliproteins found in plants, algae and cyanobacteria. Owing to the linear tetrapyrrole pigment phytochromobilin (PΦB) or phycocyanobilin (PCB) that is covalently linked to a large polypeptide via a thioether linkage, phytochromes perceive differences in the quality and quantity of light via their ability to photointerconvert between red (λmax660 nm) and far-red (λmax730 nm) light absorbing forms. Due to an efficient Z,E photoisomerization of the double bond between the C and D-ring pyrroles, phytochromes are nonfluorescent proteins with fluorescent quantum yields less than 10“3 at room temperature (Figure 1).Phytochrome genes have been cloned from a wide variety of photosynthetic organisms.

305 TRANSGENIC CLONED PIGS EXPRESSING ORANGE FLUORESCENT PROTEIN KUSABIRA-ORANGE

2008 ◽  
Vol 20 (1) ◽  
pp. 232
Author(s):  
H. Matsunari ◽  
M. Onodera ◽  
N. Tada ◽  
H. Mochizuki ◽  
E. Haruyama ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Blood Cells ◽  
Southern Blotting ◽  
Fluorescent Protein ◽  
Retroviral Vector ◽  
Fibroblast Cells ◽  
Peripheral Blood Cells ◽  
Fetal Fibroblast ◽  
Orange Fluorescent Protein ◽  
Orange Fluorescence

Transgenic (Tg)-cloned pigs expressing fluorescent proteins are very useful in research such as cell or tissue transplantation studies. In this study, we produced Tg-cloned pigs with an orange fluorescent protein, humanized Kusabira-Orange (huKO), and analyzed the characteristics of these pigs. Fetal fibroblast cells transduced with huKO gene by a gene silencing-resistant retroviral vector, pDΔNhuKO (Suzuki et al. 2002 J. Neurochem. 82, 953–960) were used as nuclear donors. Recipient cytoplasts were prepared using oocytes matured in vitro in NCSU23. The nuclear transfer (NT) embryos were transferred into oviducts of estrus-synchronized recipient gilts after culture in PZM-5 for 1 or 2 days. Cloned pigs obtained were analyzed by Southern blotting for the transgene integration. A total of 23 organs and tissues, including brain, eye, internal and reproductive organs, skin, skeletal muscle, and cartilage were examined by fluorescence stereomicroscopy. Cryo- and paraffin-embedded tissue sections were also prepared to examine fluorescence expression. Immunofluorescent staining of brain cryosection and flow cytometry analysis of peripheral blood cells were performed to identify huKO-expressing cells. Transfer of 429 NT embryos into 4 recipients resulted in 18 (4.2%) cloned offspring. Southern blotting analysis of the cloned pigs confirmed transduction of 2 to 17 copies of the huKO gene in each pig. Autopsy was performed in 6 pigs, and orange fluorescence was confirmed in all the tissues and organs examined in each pig. In addition, prominent orange fluorescence was detected in pancreatic islets and renal glomeruli, indicating that these transgenic pigs are ideal for islet cell transplantation studies. Expression of huKO was also detected in neurons, microglia, and astrocytes in the brain, and granulocytes, monocytes, macrophages, lymphocytes, and platelets in the peripheral blood cells, whereas the expression level in red blood cells was lower. Re-cloning was performed using primary culture fibroblast cells established from 2 Tg-cloned pigs. Transfer of 300 re-cloned embryos into 4 recipients resulted in 3 pregnancies. A re-cloned fetus at Day 44 of gestation showed systemic fluorescence. These results demonstrate that the retroviral vector pD?NhuKO is resistant to gene silencing in pigs, that transduction and expression of the huKO gene had no lethal effects on fetal development, and that a Tg-cloned pig expressing orange fluorescence can be produced by NT of fetal fibroblast cells transduced with the huKO gene. This study was supported by PROBRAIN.

scholarly journals Autophagy is involved in assisting the replication of Bamboo mosaic virus in Nicotiana benthamiana

2019 ◽  
Vol 70 (18) ◽  
pp. 4657-4670 ◽  
Author(s):  
Ying-Ping Huang ◽  
Ying-Wen Huang ◽  
Yung-Jen Hsiao ◽  
Siou-Cen Li ◽  
Yau-Huei Hsu ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Rna Virus ◽  
Critical Role ◽  
Viral Rna ◽  
Autophagosome Formation ◽  
Orange Fluorescent Protein ◽  
Positive Sense ◽  
Trigger Cell Death

Abstract Autophagy plays a critical role in plants under biotic stress, including the response to pathogen infection. We investigated whether autophagy-related genes (ATGs) are involved in infection with Bamboo mosaic virus (BaMV), a single-stranded positive-sense RNA virus. Initially, we observed that BaMV infection in Nicotiana benthamiana leaves upregulated the expression of ATGs but did not trigger cell death. The induction of ATGs, which possibly triggers autophagy, increased rather than diminished BaMV accumulation in the leaves, as revealed by gene knockdown and transient expression experiments. Furthermore, the inhibitor 3-methyladenine blocked autophagosome formation and the autophagy inducer rapamycin, which negatively and positively affected BaMV accumulation, respectively. Pull-down experiments with an antibody against orange fluorescent protein (OFP)-NbATG8f, an autophagosome marker protein, showed that both plus- and minus-sense BaMV RNAs could associate with NbATG8f. Confocal microscopy revealed that ATG8f-enriched vesicles possibly derived from chloroplasts contained both the BaMV viral RNA and its replicase. Thus, BaMV infection may induce the expression of ATGs possibly via autophagy to selectively engulf a portion of viral RNA-containing chloroplast. Virus-induced vesicles enriched with ATG8f could provide an alternative site for viral RNA replication or a shelter from the host silencing mechanism.

scholarly journals A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo

2016 ◽  
Vol 34 (7) ◽  
pp. 760-767 ◽  
Author(s):  
Jun Chu ◽  
Younghee Oh ◽  
Alex Sens ◽  
Niloufar Ataie ◽  
Hod Dana ◽  
...  
Keyword(s):  
Bioluminescence Imaging ◽  
Fluorescent Protein ◽  
Dual Emission ◽  
Orange Fluorescent Protein ◽  
Emission Microscopy

Characterization of Novel Orange Fluorescent Protein Cloned from Cnidarian Tube Anemone Cerianthus sp.

2007 ◽  
Vol 9 (4) ◽  
pp. 469-478 ◽  
Author(s):  
Denis Tsz-Ming Ip ◽  
Kam-Bo Wong ◽  
David Chi-Cheong Wan
Keyword(s):  
Fluorescent Protein ◽  
Orange Fluorescent Protein

scholarly journals An Orange Fluorescent Protein with a Large Stokes Shift for Single-Excitation Multicolor FCCS and FRET Imaging

2012 ◽  
Vol 134 (18) ◽  
pp. 7913-7923 ◽  
Author(s):  
Daria M. Shcherbakova ◽  
Mark A. Hink ◽  
Linda Joosen ◽  
Theodorus W. J. Gadella ◽  
Vladislav V. Verkhusha
Keyword(s):  
Fluorescent Protein ◽  
Stokes Shift ◽  
Large Stokes Shift ◽  
Orange Fluorescent Protein
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