Identification and Characterization of Potential Target Genes Critical for Doxorubicin Overproduction Using Streptomyces DNA Microarray Systems

초록

To identify and characterize the genomics-driven potential target genes critical for doxorubincin overproduction, three different types of doxorubicin overproducing strains, a dnrI (doxorubicin-specific positive regulatory gene)-overexpressor, a dnrA (gene involved in the conversion from daunarubicin to doxorubicin)-overexpressor, and a recursively-mutated industrial strain, were generated and examined their genomic transcription profiles using Streptomyces DNA microarray systems. Both the growth-phase-dependent RNA samples isolated from the wild-type and the overproduer were labeled using Cy3- and Cy5-fluorescence dyes, respectively, followed by the competitive hybridization with the doxorubicin-biosynthetic DNA chip as well as S. coelicolor whole genome chip. The DNA microarray results revealed several potential target genes in S. peucetius genome, whose expressions were significantly either up- or down-regulated comparing with the wild-type strain. A systematic understanding of doxorubicin overproduction at the genomic level presented in this research should lead us a rational design of molecular genetic strain improvement strategy.