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A Method Combining TA Cloning and Fluorescence Screening for Rapid Acquisition of Transgenic Seeds
Author(s) -
Min Li,
Lingyan Jiang,
Qin Liu,
Yuan-Hang Wu,
Guodao Liu,
Yinhua Chen,
Lijuan Luo
Publication year - 2020
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0141
Subject(s) - cloning (programming) , transformation (genetics) , transgene , biology , plasmid , vector (molecular biology) , selection (genetic algorithm) , genetically modified crops , cloning vector , microbiology and biotechnology , multiple cloning site , computational biology , genetics , gene , computer science , recombinant dna , artificial intelligence , programming language
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.

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