Region-specific transcriptional activity in the genome of Streptomyces coelicolor A3(2).
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.
Gust
B
B
Genome Research Unit, Department of Genetics, Kaiserslautern University, 67663 Kaiserslautern, Germany.
Spatz
K
K
Spychaj
A
A
Redenbach
M
M
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003360:Cosmids / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D018076:DNA, Complementary; D012329:RNA, Bacterial; D012333:RNA, Messenger