Genomics-guided technologies including cDNA microarrays have been successfully applied for the identification of gene expression alterations in various Streptomyces strains. To detect global changes in mRNA abundance associated with overproduction of anti-cancer doxorubicin, reverse engineering strategy via comparative transcriptome analyses in cultures of the wild-type and industrial mutant strains of S. peucetius were conducted using interspecies S. coelicolor cDNA microarrays. Among 160 S. coelicolor potential candidate genes showing at least a two-fold change in transcription profile, a SCO3579 (whiB-like gene named wblA) inhibited the biosynthesis of doxorubicin in S. peucetius as well as the production of antibiotics in S. coelicolor, suggesting that wblA and its homologs act globally among streptomycetes as down-regulators of antibiotic biosynthesis. In addition, one of tetR-family transcriptional regulatory genes was also identified to encode a global antibiotic regulator. Gene disruption of S. coelicolor revealed that this tetR-family regulatory gene down-regulates antibiotic biosynthesis through pathway-specific regulators even in the presence of the wblA transcript, implying that it might encode a wblA-bypassing antibiotic down-regulator. To isolate and manipulate a wblA homolog in a doxorubicin-overproducing S. peucetius industrial stain, a total genomic DNA library from a S. peucetius industrial mutant was constructed and screened using a S. coelicolor wblA as a probe, resulting in isolation of a wblA ortholog (wblA-spe) showing 95% amino acid identity to a S. coelicolor wblA. Gene disruption of wblA-spe from the S. peucetius industrial mutant resulted in additional increase of doxorubicin production, implying that doxorubicin productivity in the S. peucetius industrial mutant strain could be further improved via rational comparative transcriptomics-guided target gene manipulation.
저자
Eung-Soo Kim [ Department of Biological Engineering, Inha University ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
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