Generation of a selectable marker free, highly expressed single copy locus as landing pad for transgene stacking in sugarcane.
FLP
FLPe
Insulator
Selection marker removal
Site-specific recombination
Sugarcane
Journal
Plant molecular biology
ISSN: 1573-5028
Titre abrégé: Plant Mol Biol
Pays: Netherlands
ID NLM: 9106343
Informations de publication
Date de publication:
Jun 2019
Jun 2019
Historique:
received:
07
09
2018
accepted:
15
03
2019
pubmed:
29
3
2019
medline:
8
6
2019
entrez:
29
3
2019
Statut:
ppublish
Résumé
A selectable marker free, highly expressed single copy locus flanked by insulators was created as landing pad for transgene stacking in sugarcane. These events displayed superior transgene expression compared to single-copy transgenic lines lacking insulators. Excision of the selectable marker gene from transgenic sugarcane lines was supported by FLPe/FRT site-specific recombination. Sugarcane, a tropical C4 grass in the genus Saccharum (Poaceae), accounts for nearly 80% of sugar produced worldwide and is also an important feedstock for biofuel production. Generating transgenic sugarcane with predictable and stable transgene expression is critical for crop improvement. In this study, we generated a highly expressed single copy locus as landing pad for transgene stacking. Transgenic sugarcane lines with stable integration of a single copy nptII expression cassette flanked by insulators supported higher transgene expression along with reduced line to line variation when compared to single copy events without insulators by NPTII ELISA analysis. Subsequently, the nptII selectable marker gene was efficiently excised from the sugarcane genome by the FLPe/FRT site-specific recombination system to create selectable marker free plants. This study provides valuable resources for future gene stacking using site-specific recombination or genome editing tools.
Identifiants
pubmed: 30919152
doi: 10.1007/s11103-019-00856-4
pii: 10.1007/s11103-019-00856-4
doi:
Substances chimiques
Biofuels
0
Genetic Markers
0
Plant Proteins
0
Kanamycin Kinase
EC 2.7.1.95
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
247-263Subventions
Organisme : Office of Science
ID : DE-SC0018420
Organisme : Syngenta Crop Protection, LLC.
ID : 00108472
Organisme : CPBR
ID : 00108472
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