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A method combining TA cloning and fluorescence screening for rapid acquisition of transgenic seeds.
- Source :
-
BioTechniques [Biotechniques] 2020 May; Vol. 68 (5), pp. 251-256. Date of Electronic Publication: 2020 Mar 06. - Publication Year :
- 2020
-
Abstract
- The establishment of transgenic plants has greatly promoted the progress of plant research. However, traditional selection methods using antibiotics or herbicides may miss any positive transformants with growth defects. Additionally, screening with antibiotics/herbicides requires a huge amount of seeds, sterile work conditions and a large amount of space to germinate plants, making the selection process time- and labor-consuming. In this study, we constructed a novel stable transformation vector, plasmid of OLE1-GFP T-DNA vector (pOGT), which can shorten the steps of cloning foreign genes into expression vectors by using TA cloning. Additionally, selection of transformed seeds with fluorescence overcomes the difficulties of conventional selection with antibiotics/herbicides and simplifies the screening process for transgenic plants.
- Subjects :
- Arabidopsis classification
Arabidopsis genetics
Genetic Vectors genetics
Membrane Proteins genetics
Plant Proteins genetics
Plasmids genetics
Cloning, Molecular methods
Genes, Plant genetics
Plants, Genetically Modified classification
Plants, Genetically Modified genetics
Seeds classification
Seeds genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-9818
- Volume :
- 68
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- BioTechniques
- Publication Type :
- Academic Journal
- Accession number :
- 32141763
- Full Text :
- https://doi.org/10.2144/btn-2019-0141