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AUTHOR |
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Jeon J-S, Lee S, Jung K-H, Jun S-H, Jeong D-H, Lee J-W, Kim C, Jang S, Lee S-Y, Yang K, Nam J, An K, Han M-J, Sung R-J, Choi H-S, Yu J-H, Choi J-H, Cho S-Y, Cha S-S, Kim S-I, An G. (2000) |
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TITLE |
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T-DNA insertional mutagenesis for functional genomics in rice. |
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JOURNAL |
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Plant Journal, 22, 561-570. |
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The Abstract Of The Paper |
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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 beta-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 flowers from 7026 lines, and developing seeds from 1948 lines. The data revealed that 1.6-2.1% of tested organs were GUS-positive in the tested organs, and that their GUS expression patterns were organ- or tissue-specific or ubiquitous in all parts of the plant. The large population of T-DNA-tagged lines will be useful for identifying insertional mutants in various genes and for discovering new genes in rice.
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Copyright 2004 (c) Plant Functional Genomics Lab. Department of Biological Sciences, Kyung Hee University
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