In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals.

Journal of Bacteriology
M J CasadabanS N Cohen

Abstract

We report the construction and use of a series of plasmid vectors suitable for the detection and cloning of translational control signals and 5' coding sequences of exogenously derived genes. In these plasmids, the first eight codons of the amino-terminal end of the lactose operon beta-galactosidase gene, lacZ, were removed, and unique BamHI, EcoRI, and SmaI (XmaI) endonuclease cleavage sites were incorporated adjacent to the eighth codon of lacZ. Introduction of deoxyribonucleic acid fragments containing appropriate regulatory signals and 5' coding sequences into such lac fusion plasmids led to the production of hybrid proteins consisting of the carboxyl-terminal segment of a beta-galactosidase remnant plus a peptide fragment that contained the amino-terminal amino acids encoded by the exogenous deoxyribonucleic acid sequence. These hybrid peptides retained beta-galactosidase enzymatic activity and yielded a Lac+ phenotype. Such hybrid proteins are useful for purifying peptide sequences encoded by exogenous deoxyribonucleic acid fragments and for studies relating the structure and function of specific peptide segments.

References

Jan 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D V GoeddelA D Riggs
Apr 14, 1978·Biochemical and Biophysical Research Communications·C P BahlS A Narang
Sep 1, 1979·Journal of Bacteriology·A SarthyJ Beckwith
Jan 1, 1979·Cold Spring Harbor Symposia on Quantitative Biology·S N CohenC P Tu
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·M J Casadaban, S N Cohen
Mar 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·T E ShenkP Berg
May 1, 1978·Journal of Bacteriology·P O'FarrellD H Gelfand
Sep 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·B PoliskyH W Boyer
Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A M Maxam, W Gilbert
Apr 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A V Fowler, I Zabin
Jan 1, 1979·Cold Spring Harbor Symposia on Quantitative Biology·J G Sutcliffe
Aug 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·J ChouS N Cohen
Oct 3, 1977·European Journal of Biochemistry·J Kania, B Müller-Hill
Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·T J SilhavyM Schwartz
Oct 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·T J SilhavyJ R Beckwith
Nov 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·K BackmanW Gilbert
Mar 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·M J Casadaban
Apr 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·K E LangleyI Zabin
Apr 15, 1975·Journal of Molecular Biology·D H MitchellJ R Beckwith
Jan 1, 1974·Methods in Enzymology·D L Kacian, S Spiegelman
Aug 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·S N CohenL Hsu
Oct 15, 1973·Biochemical and Biophysical Research Communications·Y M Kupersztoch, D R Helinski
Mar 1, 1974·Journal of Bacteriology·W S ReznikoffB Magasanik
Nov 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·S N CohenR B Helling
Jun 7, 1974·Nature·B Müller-Hill, J Kania
Jan 1, 1980·Cell·M PtashneR T Sauer
May 1, 1980·Journal of Bacteriology·J K WelplyI Zabin
Apr 1, 1980·Journal of Molecular Biology·M J Casadaban, S N Cohen

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Citations

Sep 1, 1990·Bio/technology·W C Skarnes
May 1, 1991·Molecular & General Genetics : MGG·M van de GuchteG Venema
Sep 1, 1989·Molecular & General Genetics : MGG·H P Schweizer, P Datta
Nov 1, 1989·Molecular Microbiology·B E Davidson, I Saint Girons
Mar 1, 1988·Molecular & General Genetics : MGG·J W Forster, P Strike
Aug 1, 1988·Molecular & General Genetics : MGG·T Elliott, J R Roth
Nov 1, 1986·Molecular & General Genetics : MGG·N IrinoH Nakayama
Mar 1, 1988·Molecular & General Genetics : MGG·J A VericatJ Barbé
Dec 1, 1987·Molecular & General Genetics : MGG·A BirkmannA Böck
Jul 1, 1986·Molecular & General Genetics : MGG·P GamasM Chandler
Jan 1, 1984·Molecular & General Genetics : MGG·R LawsonR Voellmy
Jan 1, 1984·Molecular & General Genetics : MGG·D J OwenJ A Shapiro
Jan 1, 1993·DNA Sequence : the Journal of DNA Sequencing and Mapping·L T StaufferG V Stauffer
Nov 10, 2000·The Journal of Biological Chemistry·I MarianovskyG Glaser

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