Literature DB >> 9461479

UGUS, a reporter for use with destabilizing N-termini.

D Zaccaria Roberta Greco1, H MacWilliams, S Bozzaro, A Ceccarelli.   

Abstract

Due to constraints in vector construction, reporter polypeptides often carry N-terminal sequences of extraneous origin. Since protein half-life can be influenced by small determinants in the N-terminus, such foreign sequences can destabilize proteins and compromise results of reporter-based studies. We provide a real-life example of this problem (destabilizing sequences derived from a ribosomal protein) and show that it can be solved with the ubiquitin fusion technique, in which ubiquitin sequences are placed upstream of the reporter, in our case beta-glucuronidase. Post-translational processing by characterized pathways removes the ubiquitin together with destabilizing sequences, generating a stable reporter whose N-terminus is constant.

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Year:  1998        PMID: 9461479      PMCID: PMC147347          DOI: 10.1093/nar/26.4.1128

Source DB:  PubMed          Journal:  Nucleic Acids Res        ISSN: 0305-1048            Impact factor:   16.971


  13 in total

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Authors:  R T Baker
Journal:  Curr Opin Biotechnol       Date:  1996-10       Impact factor: 9.740

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Authors:  J W Tobias; A Varshavsky
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4.  A short amino acid sequence able to specify nuclear location.

Authors:  D Kalderon; B L Roberts; W D Richardson; A E Smith
Journal:  Cell       Date:  1984-12       Impact factor: 41.582

5.  In vivo half-life of a protein is a function of its amino-terminal residue.

Authors:  A Bachmair; D Finley; A Varshavsky
Journal:  Science       Date:  1986-10-10       Impact factor: 47.728

6.  Primary structure and regulation of vegetative specific genes of Dictyostelium discoideum.

Authors:  C K Singleton; S S Manning; R Ken
Journal:  Nucleic Acids Res       Date:  1989-12-11       Impact factor: 16.971

7.  Hygromycin resistance as a selectable marker in Dictyostelium discoideum.

Authors:  T T Egelhoff; S S Brown; D J Manstein; J A Spudich
Journal:  Mol Cell Biol       Date:  1989-05       Impact factor: 4.272

8.  Cyclin is degraded by the ubiquitin pathway.

Authors:  M Glotzer; A W Murray; M W Kirschner
Journal:  Nature       Date:  1991-01-10       Impact factor: 49.962

9.  Characterization of genes which are deactivated upon the onset of development in Dictyostelium discoideum.

Authors:  C K Singleton; R L Delude; C E McPherson
Journal:  Dev Biol       Date:  1987-02       Impact factor: 3.582

10.  DNA sequences required for expression of a Dictyostelium actin gene.

Authors:  S M Cohen; D Knecht; H F Lodish; W F Loomis
Journal:  EMBO J       Date:  1986-12-01       Impact factor: 11.598

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  1 in total

1.  Understanding the relationship between the primary structure of proteins and its propensity to be soluble on overexpression in Escherichia coli.

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Journal:  Protein Sci       Date:  2005-02-02       Impact factor: 6.725

  1 in total

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