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PMID: 6381966 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Expression of an artificial yeast TRP-gene cluster in yeast and Escherichia coli.

Molecular & general genetics : MGG ·Vol. 195 ·No. 3 ·1984-00-00 ·Pages 481-6

Niederberger P, Aebi M, Furter R, Prantl F, Hütter R

Abstract

All five tryptophan biosynthetic genes of Saccharomyces cerevisiae were unified on plasmid pME554, which is based on 2 micrometer DNA and pBR322 sequences allowing for autonomous replication in yeast and E. coli. Homologous and heterologous expression of this artificial yeast TRP-gene cluster was studied. Plasmid pME554 allowed for nearly normal growth of a yeast strain bearing auxotrophic mutations in all five TRP-genes. The plasmid-borne genes TRP2 to TRP5 were expressed and regulated normally in the frame of the general control. Gene TRP1, carried on an EcoRI/Bg/II fragment lacking the ARS1 function, was expressed poorly and did not respond to the general control like the chromosomally-borne TRP1 gene. Plasmid pME554 allowed for poor growth of E. coli strain W3110 tna- delta trpEA2 on minimal medium. Marked stimulation was observed, however, when anthranilic acid or indole were added. Accordingly, poor expression of the first Trp-enzyme anthranilate synthase and the last enzyme tryptophan synthase was found, whereas the other three genes were moderately well expressed in E. coli.

MeSH Terms
Base Sequence Cloning, Molecular DNA, Bacterial/genetics DNA, Fungal/genetics Escherichia coli/genetics,metabolism Gene Expression Regulation Genes, Bacterial Genes, Fungal Operon Plasmids Saccharomyces cerevisiae/genetics,metabolism Tryptophan/biosynthesis,genetics
Chemicals
DNA, Bacterial DNA, Fungal Tryptophan
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Niederberger P
Aebi M
Furter R
Prantl F
Hütter R
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00341450
Published
1984-00-00
Pages
481-6
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00341450
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