원문정보
초록
영어
Now that the genomes of many organisms have been sequenced and the products of newly
identified genes have been annotated, the next goal is to engineer the desired phenotypes
of organisms at will. For the phenotypic engineering of microorganisms, we have developed
novel artificial transcription factors (ATFs) capable of reprogramming innate gene expression
circuits in Escherichia coli. These ATFs are zinc finger (ZF) DNA binding proteins, with
distinct DNA-binding specificities, fused to an E. coli cyclic AMP receptor protein (CRP). By
randomly assembling 30 different types of ZFs, we have constructed more than 2.7 x 104
ATFs by combining DNA binding domains composed of three ZFs and CRP as an effector
domain. Using the resulting ATFs, we induced various phenotypic changes in E. coli and
selected for industrially important traits, such as the resistance to heat shock, osmotic pressure, and cold shock. The genes associated with the selected phenotypes were identified and characterized. These results and the general applicability of this platform clearly indicate that novel ATFs can be used as a powerful tool for the phenotypic engineering and functional genomics of microorganisms.