Protein subcellular localization assays using split fluorescent proteins
Abstract
The invention provides protein subcellular localization assays using split fluorescent protein systems. The assays are conducted in living cells, do not require fixation and washing steps inherent in existing immunostaining and related techniques, and permit rapid, non-invasive, direct visualization of protein localization in living cells. The split fluorescent protein systems used in the practice of the invention generally comprise two or more self-complementing fragments of a fluorescent protein, such as GFP, wherein one or more of the fragments correspond to one or more beta-strand microdomains and are used to "tag" proteins of interest, and a complementary "assay" fragment of the fluorescent protein. Either or both of the fragments may be functionalized with a subcellular targeting sequence enabling it to be expressed in or directed to a particular subcellular compartment (i.e., the nucleus).
- Inventors:
-
- Santa Fe, NM
- Los Alamos, NM
- Publication Date:
- Research Org.:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 971623
- Patent Number(s):
- 7,585,636
- Application Number:
- 11/295,374
- Assignee:
- Los Alamos National Security, LLC (Los Alamos, NM)
- DOE Contract Number:
- W-7405-ENG-36
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Waldo, Geoffrey S, and Cabantous, Stephanie. Protein subcellular localization assays using split fluorescent proteins. United States: N. p., 2009.
Web.
Waldo, Geoffrey S, & Cabantous, Stephanie. Protein subcellular localization assays using split fluorescent proteins. United States.
Waldo, Geoffrey S, and Cabantous, Stephanie. 2009.
"Protein subcellular localization assays using split fluorescent proteins". United States. https://www.osti.gov/servlets/purl/971623.
@article{osti_971623,
title = {Protein subcellular localization assays using split fluorescent proteins},
author = {Waldo, Geoffrey S and Cabantous, Stephanie},
abstractNote = {The invention provides protein subcellular localization assays using split fluorescent protein systems. The assays are conducted in living cells, do not require fixation and washing steps inherent in existing immunostaining and related techniques, and permit rapid, non-invasive, direct visualization of protein localization in living cells. The split fluorescent protein systems used in the practice of the invention generally comprise two or more self-complementing fragments of a fluorescent protein, such as GFP, wherein one or more of the fragments correspond to one or more beta-strand microdomains and are used to "tag" proteins of interest, and a complementary "assay" fragment of the fluorescent protein. Either or both of the fragments may be functionalized with a subcellular targeting sequence enabling it to be expressed in or directed to a particular subcellular compartment (i.e., the nucleus).},
doi = {},
url = {https://www.osti.gov/biblio/971623},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Sep 08 00:00:00 EDT 2009},
month = {Tue Sep 08 00:00:00 EDT 2009}
}
Works referenced in this record:
Protein-protein interaction of FHL3 with FHL2 and visualization of their interaction by green fluorescent proteins (GFP) two-fusion fluorescence resonance energy transfer (FRET)
journal, January 2000
- Li, Hoi Yeung; Ng, Enders Kai On; Lee, Simon Ming Yuen
- Journal of Cellular Biochemistry, Vol. 80, Issue 3, p. 293-303
Combinatorial Marking of Cells and Organelles with Reconstituted Fluorescent Proteins
journal, October 2004
- Zhang, Shifang; Ma, Charles; Chalfie, Martin
- Cell, Vol. 119, Issue 1, p. 137-144
Nuclear Localization of the ORF2 Protein Encoded by Porcine Circovirus Type 2
journal, June 2001
- Liu, Qiang; Tikoo, Suresh K.; Babiuk, Lorne A.
- Virology, Vol. 285, Issue 1
Seeing what was unseen: New analytical methods for molecular imaging
journal, January 2003
- Umezawa, Yoshio
- The Chemical Record, Vol. 3, Issue 1
A C-terminal signal prevents secretion of luminal ER proteins
journal, March 1987
- Munro, Sean; Pelham, Hugh R. B.
- Cell, Vol. 48, Issue 5
Near identity of HeLa cell galactosyltransferase with the human placental enzyme
journal, January 1990
- Watzele, G.; Berger, E. G.
- Nucleic Acids Research, Vol. 18, Issue 23
Golgi Retention Mechanism of -1,4-Galactosyltransferase: MEMBRANE-SPANNING DOMAIN-DEPENDENT HOMODIMERIZATION AND ASSOCIATION WITH α- AND β-TUBULINS
journal, May 1995
- Yamaguchi, Naoto; Fukuda, Michiko N.
- Journal of Biological Chemistry, Vol. 270, Issue 20
A short amino acid sequence able to specify nuclear location
journal, December 1984
- Kalderon, Daniel; Roberts, Bruce L.; Richardson, William D.
- Cell, Vol. 39, Issue 3, p. 499-509
Differential Importin-α Recognition and Nuclear Transport by Nuclear Localization Signals within the High-Mobility-Group DNA Binding Domains of Lymphoid Enhancer Factor 1 and T-Cell Factor 1
journal, August 1998
- Prieve, Mary G.; Guttridge, Katherine L.; Munguia, Jesus
- Molecular and Cellular Biology, Vol. 18, Issue 8
A Fluorescent Indicator for Detecting Protein−Protein Interactions in Vivo Based on Protein Splicing
journal, November 2000
- Ozawa, Takeaki; Nogami, Satoru; Sato, Moritoshi
- Analytical Chemistry, Vol. 72, Issue 21
Visualization of Interactions among bZIP and Rel Family Proteins in Living Cells Using Bimolecular Fluorescence Complementation
journal, April 2002
- Hu, Chang-Deng; Chinenov, Yurii; Kerppola, Tom K.
- Molecular Cell, Vol. 9, Issue 4, p. 789-798
Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal
journal, August 1986
- Lanford, Robert E.; Kanda, Patrick; Kennedy, Ronald C.
- Cell, Vol. 46, Issue 4
Antiparallel Leucine Zipper-Directed Protein Reassembly: Application to the Green Fluorescent Protein
journal, June 2000
- Ghosh, Indraneel; Hamilton, Andrew D.; Regan, Lynne
- Journal of the American Chemical Society, Vol. 122, Issue 23