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A GAL 4‐based targeted activation tagging system in A rabidopsis thaliana
Author(s) -
Waki Takamitsu,
Miyashima Shunsuke,
Nakanishi Miyako,
Ikeda Yoichi,
Hashimoto Takashi,
Nakajima Keiji
Publication year - 2013
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12049
Subject(s) - biology , gene , enhancer , genetics , transcription factor , mutant , arabidopsis thaliana , arabidopsis , computational biology , microbiology and biotechnology
Summary Activation tagging is a powerful tool for discovering novel genes that are not easily identified by loss‐of‐function (lof) screening due to genetic redundancy or lethality. Although the current activation tagging system, which involves a viral enhancer sequence, has been used for a decade, alternative methods that allow organ‐ or tissue‐specific activation are required to identify genes whose strong activation leads to loss of fertility or viability. Here, we established a GAL 4/ UAS activation‐tagging system in A rabidopsis thaliana . Host plants that express a synthetic transcription activator GAL 4: VP 16 ( GV ) in an organ‐ or tissue‐specific manner were transformed with a T‐ DNA harboring tandem copies of UAS , a GAL 4‐binding sequence. Using a post‐embryonic and root‐specific GV ‐expressing line as the host plant, we isolated several dominant mutants with abnormal root tissue patterns, designated as uas‐tagged root patterning ( urp ) mutants, and identified their causal genes. Notably, most URP genes encoded putative transcription factors, indicating that the GAL 4/ UAS activation tagging system effectively identifies genes with regulatory functions. lof phenotypes of most URP genes were either local patterning defects or visible only if homologous genes were disrupted simultaneously or independently. Systemic overexpression of some URP genes resulted in seedling lethality. These results indicate that GAL 4/ UAS activation tagging is a powerful method for identifying genes with biological functions that are not readily identified by conventional screening methods.