Auto-excision of selectable marker genes from transgenic tobacco via a stress inducible FLP/FRT site-specific recombination system

• Published in: Transgenic research Vol. 18; no. 3; pp. 455 - 465
• Main Authors: Woo, Hee-Jong, Cho, Hyun-Suk, Lim, Sun-Hyung, Shin, Kong-Sik, Lee, Si-Myung, Lee, Ki-Jong, Kim, Dong-Hern, Cho, Yong-Gu
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.06.2009; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: Aminobutyrates - pharmacology; Animal Genetics and Genomics; Anti-Bacterial Agents - pharmacology; Antibiotic resistance; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Cinnamates - pharmacology; Drug Resistance, Bacterial - genetics; FLP gene; Fundamental and applied biological sciences. Psychology; Genetic Engineering; Genetic technics; Genetic Vectors; Hydrogen peroxide; Hydrogen Peroxide - pharmacology; Hygromycin; Hygromycin B - analogs & derivatives; Hygromycin B - pharmacology; Life Sciences; Media (selective); Methods. Procedures. Technologies; Molecular Medicine; Nicotiana - genetics; Nicotiana - growth & development; Nicotiana - metabolism; Original Paper; Oxidative Stress; Plant Genetics and Genomics; Plants, Genetically Modified - genetics; Plants, Genetically Modified - growth & development; Plants, Genetically Modified - metabolism; Promoter Regions, Genetic; recombinase; Recombinases - genetics; Recombination; Recombination, Genetic; Shoots; Stress; Tobacco; Transformation; Transgenic animals and transgenic plants; Transgenic plants; Transgenics; Transgenic tobacco; Auto-excision; Oxidative stress-inducible; Marker-free; FLP/FRT recombination system; Oxidative stress; Recombination; Nicotiana tabacum; Site specificity; Reporter gene; Dicotyledones; Angiospermae; Spermatophyta; Transgenic plant; Solanaceae
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-008-9236-x

Antibiotic resistance marker genes are powerful selection tools for use in plant transformation processes. However, once transformation is accomplished, the presence of these resistance genes is no longer necessary and can even be undesirable. We herein describe the successful excision of antibiotic resistance genes from transgenic plants via the use of an oxidative stress-inducible FLP gene. FLP encodes a recombinase that can eliminate FLP and hpt selection genes flanked by two FRT sites. During a transformation procedure in tobacco, transformants were obtained by selection on hygromycin media. Regenerants of the initial transformants were screened for selective marker excision in hydrogen peroxide supplemented media and both the FLP and hpt genes were found to have been eliminated. About 13-41% of regenerated shoots on hydrogen peroxide media were marker-free. This auto-excision system, mediated by the oxidative stress-inducible FLP/FRT system to eliminate a selectable marker gene can be very readily adopted and used to efficiently generate marker-free transgenic plants.