Simple identification of transgenic Arabidopsis plants carrying a single copy of the integrated gene

Bioscience, Biotechnology and Biochemistry

Volumn 70, Issue 7, 2006, Pages 1780-1783

  Kihara, Tomonori ^a   Zhao, Cheng Ri ^b   Kobayashi, Yuriko ^b   Takita, Eiji ^c   Kawazu, Tetsu ^a   Koyama, Hiroyuki ^b  

^a Forestry Research Institute   (Japan)

^b GIFU UNIVERSITY   (Japan)

^c NARA INSTITUTE OF SCIENCE AND TECHNOLOGY   (Japan)

Author keywords

Arabidopsis thaliana; Genomic polymerase chain reaction (PCR); Integrated gene copy number; Transgenic plant

Indexed keywords

DNA; ENZYME KINETICS; GENES;

ARABIDOPSIS THALIANA; GENOMIC POLYMERASE CHAIN REACTION (PCR); INTEGRATED GENE COPY NUMBER; TRANSGENIC PLANT;

PLANTS (BOTANY);

PLANT DNA;

ARABIDOPSIS; ARTICLE; GENE DOSAGE; GENETICS; GENOME; HETEROZYGOTE; HOMOZYGOTE; POLYMERASE CHAIN REACTION; TRANSGENIC PLANT;

ARABIDOPSIS; DNA, PLANT; GENE DOSAGE; GENOME, PLANT; HETEROZYGOTE; HOMOZYGOTE; PLANTS, GENETICALLY MODIFIED; POLYMERASE CHAIN REACTION;

ARABIDOPSIS; ARABIDOPSIS THALIANA;

EID: 33746396985     PISSN: 09168451     EISSN: 13476947     Source Type: Journal    
DOI: 10.1271/bbb.50687     Document Type: Article

References (9)

1. Grevelding, C., Fantes, V., Kemper, E., Schell, J., and Masterson, R., Single-copy T-DNA insertions in Arabidopsis are the predominant form of integration in root-derived transgenics, whereas multiple insertions are found in leaf discs. Plant Mol. Biol., 23, 847-860 (1993).
2. Lechtenberg, B., Schubert, D., Forsbach, A., Gils, M., and Schmidt, R., Neither inverted repeat T-DNA configurations nor arrangements of tandemly repeated transgenes are sufficient to trigger transgene silencing. Plant J., 34, 507-517 (2003).
3. Mason, G., Provero, P., Vaira, A. M., and Accotto, G. P., Estimating the number of integrations in transformed plants by quantitative real-time PCR. BMC Biotechnol., 2, 20 (2002).
4. Bubner, B., Gase, K., and Baldwin, I. T., Two-fold differences are the detection limit for determining transgene copy numbers in plants by real-time PCR. BMC Biotechnol., 4, 14 (2004).
5. Honda, M., Muramoto, Y., Kuzuguchi, T., Sawano, S., Machida, M., and Koyama, H., Determination of gene copy number and genotype of transgenic Arabidopsis thaliana by competitive PCR. J. Exp. Bot., 53, 1515-1520 (2002).
6. Garcia, I., Rodgers, M., Pepin, R., Hsieh, T. F., and Matringe, M., Characterization and subcellular compartmentation of recombinant 4-hydroxyphenylpyruvate dioxygenase from Arabidopsis in transgenic tobacco. Plant Physiol., 119, 1507-1516 (1999).
7. Knight, J. S., Duckett, C. M., Sullivan, J. A., Walker, A. R., and Gray, J. C., Tissue-specific, light-regulated and plastid-regulated expression of the single-copy nuclear gene encoding the chloroplast rieske FeS protein of Arabidopsis thaliana. Plant Cell Physiol., 43, 522-531 (2002).
8. Koyama, H., Kawamura, A., Kihara, T., Hara, T., Takita, E., and Shibata, D., Overexpression of mitochondrial citrate synthase in Arabidopsis thaliana improved growth on a phosphorus-limited soil. Plant Cell Physiol., 41, 1030-1037 (2000).
9. Siebert, P. D., Chenchik, A., Kellogg, D. E., Kukyanova, K. A., and Lukyano, S. A., An improved PCR method for walking in uncloned genomic DNA. Nucl. Acids Res., 23, 1087-1088 (1995).