| Reference |
Terada R, Urawa H, Inagaki Y, Tsugane K, Iida S.
Efficient gene targeting by homologous recombination in rice.
Nat Biotechnol. 2002 Oct20(10):1030-4. (2) Terada R, Johzuka-Hisatomi Y, Saitoh M, Asao H, Iida S.
Gene targeting by homologous recombination as a biotechnological tool for rice functional genomics.
Plant Physiol. 2007 Jun144(2):846-56.(3) Yamauchi T, Johzuka-Hisatomi Y, Fukada-Tanaka S, Terada R, Nakamura I, Iida S.
Homologous recombination-mediated knock-in targeting of the MET1a gene for a maintenance DNA methyltransferase reproducibly reveals dosage-dependent spatiotemporal gene expression in rice. Plant J. 2009 Oct60(2):386-96PubMed ID12219079, 17449652, 19519802
|
| Method |
(1) Researchers describe an efficient and reproducible procedure with a strong positive/negative selection for gene targeting in rice, which feeds more than half of the world's population and is an important model plant. (2) Researchers describe an improved GT (gene targeting) procedure whereby they obtained nine independent transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a frequency of approximately 2% per surviving callus and subsequently isolated eight fertile transgenic plants without the concomitant occurrence of undesirable ectopic events, even though the rice genome carries four Adh genes, including a newly characterized Adh3 gene, and a copy of highly repetitive retroelements is present adjacent to the Adh2 gene. (3) By employing a reproducible gene-targeting procedure with positive-negative selection in rice, researchers were able to obtain fertile transgenic knock-in plants with the promoterless GUS reporter gene encoding beta-glucuronidase fused with the endogenous promoter of MET1a, one of two rice MET1 genes encoding a maintenance DNA methyltransferase. |