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TECHNICAL ADVANCE T-DNA insertional mutagenesis for functional genomics in

T-DNA insertional mutagenesis for functional genomics in
Jong-Seong Jeon, Sichul Lee, Ki-Hong Jung, Sung-Hoon Jun, Dong-Hoon Jeong, Jinwon Lee, Chanhong Kim,
Seonghoe Jang, Shinyoung Lee, Kiyoung Yang, Jongmin Nam, Kyungsook An, Min-Jung Han, Ryo-Jin Sung,
Hyun-Sook Choi, Jung-Hwa Yu, Jung-Hwan Choi, Se-Yu Cho, Sang-Su Cha, Shi-In Kim and Gynheung An*
National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Pohang University
of Science and Technology (POSTECH), Pohang 790-784, Republic of Korea
Received 14 January 2000; revised 21 March 2000; accepted 21 March 2000.
*For correspondence (fax +82 562 279 2199; e-mail genean@postech.ac.kr).
We have produced 22 090 primary transgenic rice plants that carry a T-DNA insertion, which has resulted
in 18 358 fertile lines. Genomic DNA gel-blot and PCR analyses have shown that approximately 65% of
the population contains more than one copy of the inserted T-DNA. Hygromycin resistance tests
revealed that transgenic plants contain an average of 1.4 loci of T-DNA inserts. Therefore, it can be
estimated that approximately 25 700 taggings have been generated. The binary vector used in the
insertion contained the promoterless b-glucuronidase (GUS) reporter gene with an intron and multiple
splicing donors and acceptors immediately next to the right border. Therefore, this gene trap vector is
able to detect a gene fusion between GUS and an endogenous gene, which is tagged by T-DNA.
Histochemical GUS assays were carried out in the leaves and roots from 5353 lines, mature Żowers from


Source: An, Gynheung - Department of Life Science, Pohang University of Science and Technology


Collections: Biotechnology; Biology and Medicine