skip to main content
DOE Patents title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Method For Determining And Modifying Protein/Peptide Solubilty

Abstract

A solubility reporter for measuring a protein's solubility in vivo or in vitro is described. The reporter, which can be used in a single living cell, gives a specific signal suitable for determining whether the cell bears a soluble version of the protein of interest. A pool of random mutants of an arbitrary protein, generated using error-prone in vitro recombination, may also be screened for more soluble versions using the reporter, and these versions may be recombined to yield variants having further-enhanced solubility. The method of the present invention includes "irrational" (random mutagenesis) methods, which do not require a priori knowledge of the three-dimensional structure of the protein of interest. Multiple sequences of mutation/genetic recombination and selection for improved solubility are demonstrated to yield versions of the protein which display enhanced solubility.

Inventors:
 [1]
  1. Espanola, NM
Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
OSTI Identifier:
879816
Patent Number(s):
6867042
Application Number:
10/217995
Assignee:
The Regents of the University of California (Los Alamos, NM)
Patent Classifications (CPCs):
C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English

Citation Formats

Waldo, Geoffrey S. Method For Determining And Modifying Protein/Peptide Solubilty. United States: N. p., 2005. Web.
Waldo, Geoffrey S. Method For Determining And Modifying Protein/Peptide Solubilty. United States.
Waldo, Geoffrey S. Tue . "Method For Determining And Modifying Protein/Peptide Solubilty". United States. https://www.osti.gov/servlets/purl/879816.
@article{osti_879816,
title = {Method For Determining And Modifying Protein/Peptide Solubilty},
author = {Waldo, Geoffrey S},
abstractNote = {A solubility reporter for measuring a protein's solubility in vivo or in vitro is described. The reporter, which can be used in a single living cell, gives a specific signal suitable for determining whether the cell bears a soluble version of the protein of interest. A pool of random mutants of an arbitrary protein, generated using error-prone in vitro recombination, may also be screened for more soluble versions using the reporter, and these versions may be recombined to yield variants having further-enhanced solubility. The method of the present invention includes "irrational" (random mutagenesis) methods, which do not require a priori knowledge of the three-dimensional structure of the protein of interest. Multiple sequences of mutation/genetic recombination and selection for improved solubility are demonstrated to yield versions of the protein which display enhanced solubility.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2005},
month = {3}
}

Patent:

Save / Share:

Works referenced in this record:

Strategies for thein vitroevolution of protein function enzyme evolution by random recombination of improved sequences
journal, September 1997


In Vitro Synthesis of Protein in Microbial Systems
journal, December 1973


Expression of Eukaryotic Proteins in Soluble Form inEscherichia coli
journal, March 1998


Improved green fluorescence
journal, February 1995


Rapid evolution of a protein in vitro by DNA shuffling
journal, August 1994


Rapid protein-folding assay using green fluorescent protein
journal, July 1999


Antibody scFv fragments without disulfide bonds, made by molecular evolution
journal, January 1998


Optimization of DNA shuffling for high fidelity recombination
journal, March 1997


Directed evolution of a fucosidase from a galactosidase by DNA shuffling and screening
journal, April 1997


Novel green fluorescent protein (GFP) baculovirus expression vectors
journal, January 1997


Wavelength mutations and posttranslational autoxidation of green fluorescent protein.
journal, December 1994


Determination of polyprotein processing sites by amino terminal sequencing of nonstructural proteins encoded by plum pox potyvirus
journal, February 1990


Functional and nonfunctional mutations distinguished by random recombination of homologous genes
journal, July 1997


DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution.
journal, October 1994


DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution.
journal, October 1994


Functional Expression of Horseradish Peroxidase in E. coli by Directed Evolution
journal, June 1999


Redesigning Enzyme Topology by Directed Evolution
journal, March 1998


A fosmid-based genomic map and identification of 474 genes of the hyperthermophilic archaeon Pyrobaculum aerophilum
journal, February 1997


In Vivo Characterization of Mutants of the Bacteriophage f1 Gene V Protein Isolated by Saturation Mutagenesis
journal, February 1994


Expression of an antibody fragment at high levels in the bacterial cytoplasm
journal, July 1998


Improved Green Fluorescent Protein by Molecular Evolution Using DNA Shuffling
journal, March 1996


Directed evolution: Creating biocatalysts for the future
journal, December 1996


Application of a Chimeric Green Fluorescent Protein to Study Protein-Protein Interactions
journal, November 1997


The ferritins: molecular properties, iron storage function and cellular regulation
journal, July 1996


Searching Sequence Space
journal, June 1995