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Title: 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:
 [1];  [2]
  1. Santa Fe, NM
  2. 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:

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Seeing what was unseen: New analytical methods for molecular imaging
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A C-terminal signal prevents secretion of luminal ER proteins
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Near identity of HeLa cell galactosyltransferase with the human placental enzyme
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A short amino acid sequence able to specify nuclear location
journal, December 1984


A Fluorescent Indicator for Detecting Protein−Protein Interactions in Vivo Based on Protein Splicing
journal, November 2000


Visualization of Interactions among bZIP and Rel Family Proteins in Living Cells Using Bimolecular Fluorescence Complementation
journal, April 2002


Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal
journal, August 1986


Antiparallel Leucine Zipper-Directed Protein Reassembly:  Application to the Green Fluorescent Protein
journal, June 2000