ISSN:
1617-4623
Keywords:
In vivo promoter probing
;
S1 mapping
;
Protein secretion
;
R-TEM β-lactamase
;
Streptomyces R61 DD-peptidase/PBP
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary 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, down-stream of this 247 by segment, of the Streptomyces R61 DD-peptidase-encoding gene or the Escherichia coli R-TEM β-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 β-lactamase.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00315803
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