Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host.

Détails

ID Serval
serval:BIB_B317A3568A08
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host.
Périodique
Molecular and General Genetics
Auteur⸱e⸱s
Jeenes D.J., Soldati L., Baur H., Watson J.M., Mercenier A., Reimmann C., Leisinger T., Haas D.
ISSN
0026-8925 (Print)
ISSN-L
0026-8925
Statut éditorial
Publié
Date de publication
1986
Volume
203
Numéro
3
Pages
421-429
Langue
anglais
Résumé
We examine the expression of constitutive or repressible, monocistronic genes from Pseudomonas aeruginosa and Escherichia coli after their transfer to the heterologous host. To this end, chromosomal DNA from P. aeruginosa was cloned into the mobilizable broad-host-range vector pKT240; recombinant plasmids carrying the argA, argF, or proC genes were identified by complementation of the corresponding auxotrophic mutations. The isofunctional E. coli genes and the E. coli proB gene were subcloned into pKT240 from existing recombinant plasmids. The enzyme expression specified by the Pseudomonas genes in E. coli, calculated per gene copy, ranged from 0.3%-5% of the levels observed in Pseudomonas. Fusion of the P. aeruginosa proC gene to the E. coli consensus tac promoter resulted in very high proC enzyme production in E. coli, indicating that, at least in this case, the expression barrier is essentially at the level of transcriptional initiation. The E. coli argA and argF enzymes, which are controlled by repression in their native host, were synthesized constitutively in P. aeruginosa at 5% of the levels measured in E. coli under derepressed conditions. The constitutive E. coli proB and proC genes were expressed at high levels (ca. 50%) in the heterologous host. These results support the idea that P. aeruginosa may be a more permissive host than E. coli for the heterologous expression of genes from gram-negative bacteria.
Mots-clé
Chromosomes, Bacterial/physiology, DNA Restriction Enzymes, DNA, Bacterial/isolation & purification, DNA, Recombinant/metabolism, Escherichia coli/genetics, Genes, Bacterial, Genotype, Plasmids, Promoter Regions, Genetic, Pseudomonas aeruginosa/genetics, Species Specificity
Pubmed
Web of science
Création de la notice
24/01/2008 15:00
Dernière modification de la notice
20/08/2019 16:21
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