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Title: High density growth of T7 expression strains with auto-induction option

Abstract

Disclosed is a method for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise, the transcription being under the control of a promoter whose activity can be induced by an exogenous inducer whose ability to induce said promoter is dependent on the metabolic state of said bacterial cells. Initially, a culture media is provided which includes: i) an inducer that causes induction of transcription from said promoter in said bacterial cells; and ii) a metabolite that prevents induction by said inducer, the concentration of said metabolite being adjusted so as to substantially preclude induction by said inducer in the early stages of growth of the bacterial culture, but such that said metabolite is depleted to a level that allows induction by said inducer at a later stage of growth. The culture medium is inoculated with a bacterial inoculum, the inoculum comprising bacterial cells containing cloned DNA, the transcription of which is induced by said inducer. The culture is then incubated under conditions appropriate for growth of the bacterial cells.

Inventors:
 [1]
  1. Stony Brook, NY
Issue Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1012613
Patent Number(s):
7560264
Application Number:
10/675,936
Assignee:
Brookhaven Science Associates, LLC (Upton, NY)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {
DOE Contract Number:  
AC02-98CH10886
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Studier, F William. High density growth of T7 expression strains with auto-induction option. United States: N. p., 2009. Web.
Studier, F William. High density growth of T7 expression strains with auto-induction option. United States.
Studier, F William. Tue . "High density growth of T7 expression strains with auto-induction option". United States. https://www.osti.gov/servlets/purl/1012613.
@article{osti_1012613,
title = {High density growth of T7 expression strains with auto-induction option},
author = {Studier, F William},
abstractNote = {Disclosed is a method for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise, the transcription being under the control of a promoter whose activity can be induced by an exogenous inducer whose ability to induce said promoter is dependent on the metabolic state of said bacterial cells. Initially, a culture media is provided which includes: i) an inducer that causes induction of transcription from said promoter in said bacterial cells; and ii) a metabolite that prevents induction by said inducer, the concentration of said metabolite being adjusted so as to substantially preclude induction by said inducer in the early stages of growth of the bacterial culture, but such that said metabolite is depleted to a level that allows induction by said inducer at a later stage of growth. The culture medium is inoculated with a bacterial inoculum, the inoculum comprising bacterial cells containing cloned DNA, the transcription of which is induced by said inducer. The culture is then incubated under conditions appropriate for growth of the bacterial cells.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2009},
month = {7}
}

Works referenced in this record:

The General Stress Sigma Factor ςS ofEscherichia coli Is Induced during Diauxic Shift from Glucose to Lactose
journal, January 1998


cAMP receptor protein-cAMP plays a crucial role in glucose-lactose diauxie by activating the major glucose transporter gene in Escherichia coli
journal, November 1997


Novel escherichia coli strain allows efficient recombinant protein production using lactose as inducer
journal, June 2003


Regulation of sCD4-183 gene expression from phage-T7-based vectors in Escherichia coli
journal, April 1995


Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor
journal, May 1991


Auto-induction medium for the production of [U-15N]- and [U-13C, U-15N]-labeled proteins for NMR screening and structure determination
journal, April 2005


Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes
journal, May 1986


Protein production by auto-induction in high-density shaking cultures
journal, May 2005


Protocols for production of selenomethionine-labeled proteins in 2-L polyethylene terephthalate bottles using auto-induction medium
journal, April 2005