Journal article
Cold inducible promoter driven Cre-lox system proved to be highly efficient for marker gene excision in transgenic barley
Journal of Biotechnology, Vol.265, pp.15-24
2018
Abstract
A Cre-lox based auto-excision strategy has been adapted for barley, capable of cre and selectable marker gene (SMG) removal. The cold inducible wheat promoter called wcs120 was utilised for driving Cre expression. The binary vector was carrying the transgene (uidA) and a so called ‘recombination cassette’ flanked by the lox sequences. This part included both the recombinase gene and the SMG (bar) under the control of a constitutive promoter. T0, T1 and T2 transgenic plants were subjected to low temperature (at 4 °C, 10 °C and 12 °C) at different developmental stages to induce recombination. The presence of uidA, cre, and bar genes and recombination footprints were studied by PCR and DNA sequencing, while cre transcription was followed by qRT-PCR. These analyses indicated that, cold treatment of the germinating seeds (4 °C for 3 days) followed by plant growing at higher temperature (24 °C) has been the most efficient (90–100%), and this treatment lead to heritable changes in the genome. Thermal separation of Cre accumulation (at low temperature) from Cre enzyme activity (at higher temperature) could have prevented the premature excision of its own encoding gene, and lead to high expression level thereby increasing recombination frequency.
Details
- Title
- Cold inducible promoter driven Cre-lox system proved to be highly efficient for marker gene excision in transgenic barley
- Authors/Creators
- C. Éva (Author/Creator) - Eötvös Loránd UniversityF. Téglás (Author/Creator) - Eötvös Loránd UniversityH. Zelenyánszki (Author/Creator) - Eötvös Loránd UniversityC. Tamás (Author/Creator) - Hungarian Academy of SciencesA. Juhász (Author/Creator) - Hungarian Academy of SciencesK. Mészáros (Author/Creator) - Hungarian Academy of SciencesL. Tamás (Author/Creator) - Eötvös Loránd University
- Publication Details
- Journal of Biotechnology, Vol.265, pp.15-24
- Publisher
- Elsevier BV
- Identifiers
- 991005541363807891
- Copyright
- © 2017 Elsevier B.V.
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- International collaboration
- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.4 Crop Science
- 3.4.119 Micropropagation
- Web Of Science research areas
- Biotechnology & Applied Microbiology
- ESI research areas
- Biology & Biochemistry