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Phenotypic Alteration and Target Gene Identification Using Combinatorial Libraries of Zinc Finger Proteins in Prokaryotic Cells

Kyung-Soon Park, Young-Soon Jang, Horim Lee, and Jin-Soo Kim^*

ToolGen, Inc., 461-6 Jeonmin-dong, Yuseong-gu, Daejeon, South Korea 305-390

Received 9 March 2005/ Accepted 27 April 2005

We have developed a method with prokaryotic organisms that uses randomized libraries of zinc finger-containing artificial transcription factors to induce phenotypic variations and to identify genes involved in the generation of a specific phenotype of interest. Combining chromatin immunoprecipitation experiments and in silico prediction of target DNA binding sequences for the artificial transcription factors, we identified ubiX, whose down-regulation correlates with the thermotolerance phenotype in Escherichia coli. Our results show that randomized libraries of artificial transcription factors are powerful tools for functional genomic studies.

These authors contributed equally to this work.

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