Transcriptional Analysis of the dd-Peptidase/Penicillin-Binding Protein-Encoding dac Gene of Streptomyces R61: Use of the Promoter and Signal Sequences in a Secretion Vector
In vivo promoter probing; S1 mapping; Protein secretion; R-TEM beta-lactamase; Streptomyces; R61 DD-peptidase/PBP
Abstract :
[en] The promoter region of the gene encoding the extracellular DD-peptidase/penicillin-binding protein of Streptomyces R61 has been identified by in vivo promoter probing and S1 mapping. A secretion vector, pDML116, was constructed by inserting into the multicopy Streptomyces plasmid pIJ702, a 247 bp DNA sequence that contained the transcriptional, translational and secretory signals and the 12 amino acid N-terminal region-encoding sequence of the mature Streptomyces DD-peptidase/penicillin-binding protein. Insertion, downstream of this 247 bp segment, of the Streptomyces R61 DD-peptidase-encoding gene or the Escherichia coli R-TEM beta-lactamase-encoding gene yielded plasmids pDML120 and pDML128, respectively, which allowed expression and secretion of the relevant enzymes by Streptomyces lividans. The maximal secretion levels obtained were 42 mg protein/ml for the autologous Streptomyces DD-peptidase and 0.9 mg protein/ml for the heterologous E. coli beta-lactamase.
Piron-Fraipont, Claudine; Université de Liège - ULiège > Institut de Chimie > Département de Microbiologie
Lenzini, Mauro V; Université de Liège - ULiège > Insitut de Chimie > Département de Microbiologie
Dusart, Jean; Université de Liège - ULiège > Institut de Chimie > Département de Microbiologie
Ghuysen, Jean-Marie ; Université de Liège - ULiège > Institut de Chimie > Département de Microbiologie
Language :
English
Title :
Transcriptional Analysis of the dd-Peptidase/Penicillin-Binding Protein-Encoding dac Gene of Streptomyces R61: Use of the Promoter and Signal Sequences in a Secretion Vector
Publication date :
01 August 1990
Journal title :
Molecular and General Genetics
ISSN :
0026-8925
Publisher :
Springer
Issue :
223
Pages :
114-120
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FRSM - Fonds de la Recherche Scientifique Médicale
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