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 Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chary, V. K. Articles by Martin, J. F. Search for Related Content PubMed PubMed Citation Articles by Chary, V. K. Articles by Martin, J. F. Agricola Articles by Chary, V. K. Articles by Martin, J. F.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 08 1997, 2977-2982, Vol 63, No. 8
Copyright © 1997, American Society for Microbiology

Amy as a reporter gene for promoter activity in Nocardia lactamdurans: comparison of promoters of the cephamycin cluster

VK Chary, JL de la Fuente, P Liras and JF Martin
Faculty of Biology, University of Leon, Spain.

Promoter probe vectors containing the pA origin of replication and the Streptomyces griseus promoterless amy gene (encoding alpha-amylase) as reporter have been constructed to study transcription initiation regions in Nocardia lactamdurans. In some of the promoter probe vectors the phage fd terminator has been introduced to avoid readthrough expression from upstream sequences. By using these vectors, four different transcription initiation regions of the cephamycin gene cluster have been studied in N. lactamdurans. The bla gene encoding a beta-lactamase has a relatively strong promoter. Two other separate promoters corresponding to the lat and cefD genes (encoding, respectively, lysine-6-aminotransferase and isopenicillin N-epimerase) showed weak transcription initiation ability. These two promoters are arranged in a bidirectional transcription initiation region located in the center of the cephamycin gene cluster. The cmcH gene (encoding 3- hydroxymethylcephem carbamoyltransferase) upstream region did not contain a functional promoter, suggesting that cmcH is transcribed as a part of a polycistronic mRNA. The native amy promoter is used very efficiently in N. lactamdurans, resulting in secretion of high levels of extracellular alpha-amylase.

This article has been cited by other articles:

• de la Fuente, A., Lorenzana, L. M., Martin, J. F., Liras, P. (2002). Mutants of Streptomyces clavuligerus with Disruptions in Different Genes for Clavulanic Acid Biosynthesis Produce Large Amounts of Holomycin: Possible Cross-Regulation of Two Unrelated Secondary Metabolic Pathways. J. Bacteriol. 184: 6559-6565 [Abstract] [Full Text]  
• Santamarta, I., Rodriguez-Garcia, A., Perez-Redondo, R., Martin, J. F., Liras, P. (2002). CcaR Is an Autoregulatory Protein That Binds to the ccaR and cefD-cmcI Promoters of the Cephamycin C-Clavulanic Acid Cluster in Streptomyces clavuligerus. J. Bacteriol. 184: 3106-3113 [Abstract] [Full Text]  
• Mellado, E., Lorenzana, L. M., Rodriguez-Saiz, M., Diez, B., Liras, P., Barredo, J. L. (2002). The clavulanic acid biosynthetic cluster of Streptomyces clavuligerus: genetic organization of the region upstream of the car gene. Microbiology 148: 1427-1438 [Abstract] [Full Text]  
• Pérez-Redondo, R., Rodríguez-García, A., Martín, J. F., Liras, P. (1999). Deletion of the pyc Gene Blocks Clavulanic Acid Biosynthesis Except in Glycerol-Containing Medium: Evidence for Two Different Genes in Formation of the C3 Unit. J. Bacteriol. 181: 6922-6928 [Abstract] [Full Text]