Title: Translation-coupling systems

Abstract

Disclosed are systems and methods for coupling translation of a target gene to a detectable response gene. A version of the invention includes a translation-coupling cassette. The translation-coupling cassette includes a target gene, a response gene, a response-gene translation control element, and a secondary structure-forming sequence that reversibly forms a secondary structure masking the response-gene translation control element. Masking of the response-gene translation control element inhibits translation of the response gene. Full translation of the target gene results in unfolding of the secondary structure and consequent translation of the response gene. Translation of the target gene is determined by detecting presence of the response-gene protein product. The invention further includes RNA transcripts of the translation-coupling cassettes, vectors comprising the translation-coupling cassettes, hosts comprising the translation-coupling cassettes, methods of using the translation-coupling cassettes, and gene products produced with the translation-coupling cassettes.

Inventors:

Pfleger, Brian; Mendez-Perez, Daniel

Issue Date:

2013-11-05

Research Org.:

Wisconsin Alumni Research Foundation (Madison, WI)

Sponsoring Org.:

USDOE

OSTI Identifier:

1107809

Patent Number(s):

8574898

Application Number:

12/976,606

Assignee:

Wisconsin Alumni Research Foundation (Madison, WI)

Patent Classifications (CPCs):

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

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C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N15/67 - General methods for enhancing the expression
C12N15/70 - Vectors or expression systems specially adapted for E. coli
C12N15/1086 - {Preparation or screening of expression libraries, e.g. reporter assays}

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/025 - {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N15/1075 - {by coupling phenotype to genotype, not provided for in other groups of this subclass}

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DOE Contract Number:  

FC02-07ER64494

Resource Type:

Patent

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES