IP Library › Granted Patent US 10,087,463
Granted Patent B2
US 10,087,463 · App. 11/454,947 · Granted Oct 2, 2018

Methods and compositions for transposition using minimal segments of the eukaryotic transformation vector piggyBac

Inventors: Malcolm J. Fraser (Granger, IN); Xu Li (Sharon, MA)
Assignee: University of Notre Dame Du Lac
C12N15/85C12N15/90C12N2800/204C12N2800/90
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Quick Facts
Patent No.
US 10,087,463
App. No.
11/454,947
Granted
Oct 2, 2018
Kind
B2
Abstract

The 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.

Claims (4)

1. A plasmid selected from the group consisting of pBSII-ECFP-R 4 /L 2 deposited as ATCC Accession # PTA-122185, pBSII-ECFP-R 4 /L 3 deposited as ATCC Accession # PTA-122183, and pBSII-ECFP-R 4 /L 4 deposited as ATCC Accession # PTA-122184.

2. The plasmid of claim 1 , wherein the plasmid is ECFP-R 4 /L 2 deposited as ATCC Accession # PTA-122185.

3. The plasmid of claim 1 , wherein the plasmid is pBSII-ECFP-R 4 /L 3 deposited as ATCC Accession # PTA-122183.

4. The plasmid of claim 1 wherein the plasmid is pBSII-ECFP-R 4 /L 4 deposited as ATCC Accession # PTA-122184.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 31, 2006
From: FRASER, MALCOLM J.; LI, XU
To: UNIVERSITY OF NOTRE DAME DU LAC
Reel/Frame 018460/0104 →
Continuity (5)
Continuation In Part 10826523 · Apr 19, 2004
Continuation In Part 10001189 · Oct 30, 2001
Provisional Application 60244984 · Nov 1, 2000
Provisional Application 60244667 · Oct 31, 2000
Related Publication 20100221824A1 · Sep 2, 2010
Cited By (2)
US 12,391,960 US 12,655,443