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PMID: 341150 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Production of a functional eukaryotic enzyme in Escherichia coli: cloning and expression of the yeast structural gene for imidazole-glycerolphosphate dehydratase (his3).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 74 ·No. 12 ·1977-12-00 ·Pages 5255-9

Struhl K, Davis RW

Abstract

A cloned segment of yeast DNA containing the structural gene for imidazoleglycerolphosphate dehydratase (D-erythro-imidazoleglycerolphosphate hydro-lase, EC 4.2.1.19) is transcribed and translated in Escherichia coli with sufficient fidelity to produce functional enzyme. This segment of yeast DNA was isolated as a viable molecular hybrid of bacteriophage lambda (lambdagt-Sc2601) which complements a nonrevertible hisB auxotroph of E. coli lacking dehydratase activity. The equivalent segments of DNA cloned from two independent his3 mutants of yeast lacking IGP dehydratase activity do not complement the hisB auxotroph. The two nonfunctional his3 alleles cloned in bacteriophage lambda can be recombined in E. coli to generate a hybrid phage which complements the hisB auxotroph. The dehydratase activity produced in E. coli by the cloned segment of yeast DNA strongly resembles the activity found in yeast.

MeSH Terms
DNA, Recombinant Escherichia coli/genetics Genes Glycerophosphates Histidine/biosynthesis Hydro-Lyases/genetics,metabolism Imidazoles Operon Saccharomyces cerevisiae/genetics Suppression, Genetic
Chemicals
DNA, Recombinant Glycerophosphates Imidazoles Histidine Hydro-Lyases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Struhl K
Davis R W
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.74.12.5255
Published
1977-12-00
Pages
5255-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC431671
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.74.12.5255 PMC Full Text View journal details
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