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KMID : 0613820060160030540
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Journal of Life Science
2006 Volume.16 No. 3 p.540 ~ p.545
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Generation and Selection of Promoter Trap Lines for the Investigation of Shoot Development in Arabidopsis
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Lee Hwa-Mok
Park Hee-Yeon
Lee Choon-Hwan
Ismayil S.Zulfugarov
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Abstract
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T-DNA-mediated transformation is a common method for generating transgenic plants with insertional mutagenesis. In order to identify important genes involved in shoot development, a system of promoter trap insertional mutagenesis was employed in Arabidopsis thaliana. For this system, an efficient promoter trap vector, pFGL561 was developed. The pFGL561 includes a basta-resistant gene, an intron with multiple splicing donor and acceptor sites, and a promoter-less GFP reporter gene. Using floral-dipping method, we made total 300 T₁ promoter-trap lines which were screened for GFP expression. GFP signals in the T₁ plants were detected with high frequency, 26.7%, and the signals were reconfirmed in T₂ plants. To isolate the genes that are involved in shoot development, phenotypes were analyzed in T₂ plants of the 19 T₁ lines that had GFP signals in shoot apex, and 6 T₁ lines were selected that had abnormal shoot development. These lines will be very useful for the investigation of shoot development.
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KEYWORD
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Arabidopsis, promoter trap lines, pFGL561, shoot development
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