Cloning and amplified expression in Streptomyces lividans of the gene encoding the extracellular beta-lactamase of Actinomadura R39.
By using the promoter-probe plasmid pIJ424, genomic DNA fragments of Actinomadura R39 were shown to have promoter activity in Streptomyces lividans. The same 100-200-copy-number plasmid was used to clone in S. lividans TK24, the gene that encodes the Actinomadura R39 beta-lactamase. Gene cloning resulted in an amplified expression of the beta-lactamase when compared with the amounts of enzyme produced by the original strain (1 mg versus 0.008 mg.litre of culture-1).
Piron-Fraipont
C
C
Département de Microbiologie, Université de Liège, Sart Tilman, Belgium.
Duez
C
C
Matagne
A
A
Molitor
C
C
Dusart
J
J
Frère
J M
JM
Ghuysen
J M
JM
D000192:Actinomycetales / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005784:Gene Amplification; D007700:Kinetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D001618:beta-Lactamases / Q000096:biosynthesis*
D000596:Amino Acids; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D001618:beta-Lactamases