Efficient yeast one-/two-hybrid screening using a library composed only of transcription factors in Arabidopsis thaliana.
Yeast one-hybrid screening is widely used for the identification of transcription factors (TFs) that interact with specific DNA sequences. However, screening a whole cDNA library is not efficient for the identification of TFs because TF genes represent only a small percentage of clones in a cDNA library. Here, we present the development of an efficient yeast one-hybrid screening system using a prey library composed only of approximately 1,500 TF cDNAs of Arabidopsis thaliana. This library enabled us to isolate a TF that binds to a specific promoter sequence with high efficiency, even when the promoter region of the gene of interest was directly employed as bait. Furthermore, this library was also successfully applied as a yeast two-hybrid library to find TFs that interact with specific proteins. This efficient system will contribute to the elucidation of gene regulatory networks in plants.
Mitsuda
Nobutaka
N
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, Higashi 1-1-1, Tsukuba, 305-8562 Japan. nobutaka.mitsuda@aist.go.jp
Ikeda
Miho
M
Takada
Shinobu
S
Takiguchi
Yuko
Y
Kondou
Youichi
Y
Yoshizumi
Takeshi
T
Fujita
Miki
M
Shinozaki
Kazuo
K
Matsui
Minami
M
Ohme-Takagi
Masaru
M
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D015723:Gene Library; D017930:Genes, Reporter; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D010940:Plant Proteins; D014157:Transcription Factors