Back to Search Start Over

A method combining TA cloning and fluorescence screening for rapid acquisition of transgenic seeds.

Authors :
Li M
Jiang LY
Liu Q
Wu YH
Liu GD
Chen YH
Luo LJ
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.

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