This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Krause, M Articles by Kleinkauf, H Search for Related Content PubMed PubMed Citation Articles by Krause, M Articles by Kleinkauf, H

Molecular cloning of an ornithine-activating fragment of the gramicidin S synthetase 2 gene from Bacillus brevis and its expression in Escherichia coli.

ABSTRACT

Partially digested chromosomal DNA of Bacillus brevis ATCC 9999, a producer of the cyclic peptide antibiotic gramicidin S, was ligated into the BamHI site of the Escherichia coli expression vector pUR2-Bam. The ligated molecules were used to transfer E. coli to ampicillin resistance. Of 5 X 10(3) colonies tested by in situ immunoassay for a cross-reaction with antibodies against the gramicidin S synthetase 2, 6 colonies were found to be immunoreactive. A clone designated MK2, which had a 3.9-kilobase insert of B. brevis DNA, directed in E. coli under the lac promoter control the synthesis of polypeptides that were cross-reactive with the antibody to the gramicidin S synthetase 2. Partial purification of the gene products by gel filtration revealed a major fraction with an approximate molecular weight of 140,000 and with specific ornithine-dependent ATP-32PPi and 2'-dATP-32PPi exchange activities. These unique activities of the gramicidin S synthetase 2 were not detected in the E. coli strain harboring the vector.

This article has been cited by other articles:

• Elsner, A., Engert, H., Saenger, W., Hamoen, L., Venema, G., Bernhard, F. (1997). Substrate Specificity of Hybrid Modules from Peptide Synthetases. J. Biol. Chem. 272: 4814-4819 [Abstract] [Full Text]  
• Stachelhaus, T, Schneider, A, Marahiel, M. (1995). Rational design of peptide antibiotics by targeted replacement of bacterial and fungal domains. Science 269: 69-72 [Abstract]  
• Stachelhaus, T., Marahiel, M. A. (1995). Modular Structure of Peptide Synthetases Revealed by Dissection of the Multifunctional Enzyme GrsA. J. Biol. Chem. 270: 6163-6169 [Abstract] [Full Text]