Optimized T-DNA transfer and vectors therefor
AssigneeCropDesign N.V.
InventorsAnna Depicker, Vladimir Mironov, Franky Terras, Willem Broekaert, Sylvie De Buck, Chris De Wilde
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Abstract
The present invention relates to T-DNA vectors and methods for obtaining transgenic eukaryotes using said vectors. The transgenic eukaryotes are characterized in that they contain the T-DNA but not the illegitimately transferred vector backbone sequence. This is achieved by modifying the T-DNA borders such that they are more efficiently nicked or such that they allow elimination of illegitimately transferred vector backbone sequences by means of recombination.
Claims25
A T-DNA transformation vector comprising T-DNA with flanking right and left T-DNA borders, wherein said right and left T-DNA borders are modified, wherein the modified right border comprises a single right border core sequence flanked by a right border outer region, and wherein the modified left T-DNA border comprises
- i) the inclusion of at least two left border core sequences separated by a sequence of at least 10-20 bp optionally carrying stop codons in three reading frames and in both directions, wherein the at least two left border core sequences are flanked only by a left border outer region;
- or
- ii) the inclusion of at least one integral nopaline-type left border region adjacent to and downstream or upstream of an integral octopine-type left border region.
A T-DNA transformation vector comprising T-DNA with flanking right and left T-DNA borders, wherein said right and left T-DNA borders are modified,
- wherein the modified right border comprises a single right border core sequence flanked by a right border outer region, and
- wherein the modified left T-DNA border comprises a single left border core sequence, wherein the single left border core sequence is further flanked by a natural left border outer region and an intra-T-DNA left border proximal region with a length of 10 to 100 bp, and which is enriched in the number of A- and T-residues, the percentage of AT-residues being 65 to 85%.
Dependencies are inferred from the claim text.