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Applied and Environmental Microbiology, August 2001, p. 3598-3602, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3598-3602.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Region-Specific Transcriptional Activity in the Genome of Streptomyces coelicolor A3(2)

Bertolt Gust,dagger Kerstin Spatz, Alexander Spychaj, and Matthias Redenbach*

Genome Research Unit, Department of Genetics, Kaiserslautern University, 67663 Kaiserslautern, Germany

Received 5 February 2001/Accepted 27 April 2001

Transcriptional analysis of microbial genomes is an important component of functional genomics. Strategies such as hybridization of labeled total RNA against ordered clone libraries or differential-display approaches have already been carried out to identify expressed genes. We describe here an additional method which applies subtractive hybridization between genome-specific DNA and total RNA followed by a PCR approach to identify expressed microbial genes. With the new strategy, the expression of genes in the terminal regions of the linear Streptomyces coelicolor A3(2) chromosome and the accessory linear plasmid SCP1 was analyzed. The results indicate that the method is useful for the identification of expressed genes in actinomycetes and other microbial systems. We demonstrate for the first time that at least 24 genes in the chromosome end regions (silent regions) of S. coelicolor are actively expressed. In addition, several expressed SCP1 genes were identified, including a gene which shows high similarity to microbial dnaN genes and which seems to play a role in SCP1 maintenance.

* Corresponding author. Mailing address: Genome Research Unit, Department of Genetics, Kaiserslautern University, Erwin-Schrödinger-Str., 67663 Kaiserslautern, Germany. Phone: 49 631 205 3250. Fax: 49 631 205 4090. E-mail: redenbac{at}rhrk.uni-kl.de.

dagger Present address: Department of Genetics, John Innes Centre, Norwich NR4 7UH, United Kingdom.

Applied and Environmental Microbiology, August 2001, p. 3598-3602, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3598-3602.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.




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