Phenotypic alteration and target gene identification using combinatorial libraries of zinc finger proteins in prokaryotic cells.
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.
Park
Kyung-Soon
KS
ToolGen, Inc., 461-6 Jeonmin-dong, Yuseong-gu, Daejeon, South Korea 305-390.
Jang
Young-Soon
YS
Lee
Horim
H
Kim
Jin-Soo
JS
D000595:Amino Acid Sequence; D002262:Carboxy-Lyases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D010641:Phenotype; D017434:Protein Structure, Tertiary / Q000235:genetics; D013696:Temperature; D014157:Transcription Factors; D016335:Zinc Fingers / Q000235:genetics*
D029968:Escherichia coli Proteins; D019151:Peptide Library; D014157:Transcription Factors; D002262:Carboxy-Lyases; C113304:ubiX protein, E coli