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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
Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
971623
Patent Number(s):
7585636
Application Number:
11/295,374
Assignee:
Los Alamos National Security, LLC (Los Alamos, NM)
Patent Classifications (CPCs):
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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. Tue . "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 = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2009},
month = {9}
}

Works referenced in this record:

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Nuclear Localization of the ORF2 Protein Encoded by Porcine Circovirus Type 2
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A C-terminal signal prevents secretion of luminal ER proteins
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A Fluorescent Indicator for Detecting Protein−Protein Interactions in Vivo Based on Protein Splicing
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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
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