Introduction of a mini-gene encoding a five-amino acid peptide confers erythromycin resistance on Bacillus subtilis and provides temporary erythromycin protection in Proteus mirabilis

• Published in: FEMS microbiology letters 2000-01, Vol.182 (2), p.213-218
• Main Authors: Novikova, Svetlana I., Bushueva, Anastassia M., Trachuk, Lesya A., Konstantinova, Galina E., Serkina, Anna V., Hoischen, Christian, Gumpert, Johannes, Chestukhina, Galina G., Mankin, Alexander, Shevelev, Alexei B.
• Format: Article
• Language: English
• Subjects: Acid resistance; Amino Acid Sequence; Amino acids; Anti-Bacterial Agents - pharmacology; Antibiotic resistance; Antibiotics; Bacillus - enzymology; Bacillus - genetics; Bacillus subtilis; Bacillus subtilis - drug effects; Bacillus subtilis - genetics; Bacillus subtilis - growth & development; Bacteria; Base Sequence; Drug Resistance, Microbial - genetics; E coli; Endopeptidases - genetics; Endopeptidases - metabolism; Endotoxins; Erythromycin; Erythromycin - pharmacology; Escherichia coli - drug effects; Escherichia coli - genetics; Gene expression; Genes; Gram-negative bacteria; L Forms - drug effects; L Forms - genetics; L Forms - growth & development; Markers; Microbiology; Molecular Sequence Data; Oligopeptides - genetics; Oligopeptides - pharmacology; Peptides; Plasmids; Plasmids - genetics; Protein Biosynthesis; Proteus mirabilis; Proteus mirabilis - drug effects; Proteus mirabilis - genetics; Proteus mirabilis - growth & development; Recombinant Proteins; rRNA; rRNA 23S; Serine Endopeptidases; Transcription; Transformation, Bacterial; Translation
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.2000.tb08897.x

Abstract A 15-bp mini-gene was introduced into Bacillus subtilis and into stable protoplast-like L-forms of Proteus mirabilis. This mini-gene encoded the peptide MVLFV and modeled a fragment of Escherichia coli 23S rRNA responsible for E. coli erythromycin (Ery) resistance. Expression of the introduced mini-gene conferred permanent Ery resistance on B. subtilis. In L-forms of P. mirabilis, the Ery-protective effect was maintained in the course of several generations. Herewith, the mechanism of Ery resistance mediated by expression of specific short peptides was shown to exist in evolutionary distant bacteria. Three new plasmids were constructed containing the gene under study transcriptionally fused with the genes encoding glutamylendopeptidase of Bacillus licheniformis or δ-endotoxin of Bacillus thuringiensis. The Ery resistance pentapeptide (E-peptide) mini-gene served as an efficient direct transcriptional reporter and allowed to select bacillar glutamylendopeptidase with improved productivity. The mini-genes encoding E-peptides may be applied as selective markers to transform both Gram-positive and Gram-negative bacteria. The small size of the E-peptide mini-genes makes them attractive selective markers for vector construction.