10087463OCR.pdf (12.13 MB)
Methods and compositions for transposition using minimal segments of the eukaryotic transformation vector piggyBac
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posted on 2019-02-18, 00:00 authored by Malcolm J. Fraser, Xu LiThe present invention provides a method for transforming an insect genome that has a much enhanced transformation frequency. The vectors and plasmids employed in the method are further described as transposition vectors that include a minimal amount of nucleotide sequence homologous to a 5' region and a 3' region of a native piggyBac nucleic acid sequence. The transformed cells or embryos may also be developed into transgenic organisms. Disclosed are minimal piggyBac-based plasmid constructs that comprises a minimal nucleic acid sequence homologous to a 5' end of a piggyBac nucleic acid sequence (about 60-80 bp, particularly 66 bp) and a relatively long (300 to about 380 bp, particularly 311 bp or 378 bp) continuous nucleic acid sequence homologous to a 3' end of a piggyBac native nucleic acid sequence. Methods employing these constructs include the use of a helper plasmid. Transformation frequencies employing the constructs are enhanced 100-fold or higher over that transformation frequency obtained using other than the herein described constructs.
History
Patent Number
US 10087463 B2Other Application
11/454,947Inventor
Xu Li Malcolm J. FraserInventor from Local Institution
Xu Li Malcolm J. FraserAssignee
University of Notre Dame Du LacDate Modified
2019-02-18Language
- English
Claims
4Prior Publication Number
US 20100221824 A1Publisher
U.S. Patent and Trademark OfficeCooperative Patent Classification Codes
C12N 15/85 (20130101); C12N 15/90 (20130101); C12N 2800/90 (20130101); C12N 2800/204 (20130101)Contributor
Xu Li|Malcolm J. FraserInternational Patent Classification Codes
C12N 15/85 (20060101); C12N 15/90 (20060101)Usage metrics
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