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Granted patent and patent applications filed by Pioneer Hi-Bred Int.

The United States patent discloses a co-transformation method in which an Agrobacterium strain has two binary vectors, each of them carrying a T-DNA region with a different heterologous sequence, and a helper plasmid. A plant scorable marker used for the selection of the transformed plant tissue is one of the heterologous sequences that can be present in the T-DNA region. The heterologous regions integrate into the genome of a plant in an unlinked manner, facilitating the retention of the genes of interest and the elimination of undesirable genes in the progeny.

The PCT application, also converted into European, Canadian and Australian patents, are also directed to the same topic.

Patents on modified binary vectors and methods – Granted patent and patent applications filed by Pioneer Hi-Bred Int.

Claims in plain English

Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6265638

  • Earliest priority – 1 October 1998
  • Filed – 28 September 1999
  • Granted – 24 July 2001
  • Expected expiry – 27 September 2019
 Title – Method of plant transformation

Claim 1
An Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids that are stably inherited, wherein each of said binary vector plasmids comprises at least one T-DNA region comprising a heterologous nucleotide sequence of interest.
Claim 12
A method for co-transformation of a plant with at least two unique heterologous nucleotide sequences of interest, said method comprising the steps ofa) contacting a tissue from said plant with an Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids that are stably inherited, wherein each of said binary vector plasmids comprises at least one T-DNA region, wherein each of said T-DNA regions comprises one of said unique nucleotide sequences, wherein at least one of said nucleotide sequences comprises at least an expression cassette comprising a plant scorable marker gene;
b) co-cultivating said tissue with said Agrobacterium;
c) culturing the tissue in a culture medium comprising an antibiotic capable of inhibiting the growth of Agrobacterium;
d) screening the tissue for expression of said plants scorable marker gene; and
e) regenerating a transformed plant from tissue that expresses said plant scorable marker gene.

The present United States patent claims:

  • an Agrobacterium strain having at least two binary vectors with T-DNA regions containing a heterologous sequence each (which are not necessarily genes and further, are not necessarily different sequences), and a helper plasmid; and
  • a method to co-transform a plant tissue by co-cultivating the tissue with an Agrobacterium strain as described above, where at least one of the heterologous sequences is an expression cassette comprising scorable marker and is used to screen for the transformed tissue. A transformed plant is regenerated from the selected transformed tissue.

Without specifying that limits of the heterologous sequences in claim 1, possible ambiguities arise.   For example, how large a sequence does a heterologous sequence have to be before it falls within the scope of the claim?  A single nucleotide? The claim also could cover scenarios where the two vectors each have identical heterologous sequences and so be anticipated by art to a single type of binary plasmid vector including a heterologous sequence provided with a helper plasmid.

Pioneer Hi-Bred International
EP 1117816

  • Earliest priority – 1 October 1998
  • Filed – 28 September 1999
  • Granted – 21 December 2005
  • Expected expiry – 27 September 2019
 Title – Method of plant transformation

Claim 1
An Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region comprising a heterologous nucleotide sequence of interest.
Claim 12
A method for co-transformation of a plant with at least two unique heterologous nucleotide sequences of interest, said method comprising the steps of:a) contacting a tissue from said plant with an Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region, wherein each of said T-DNA regions comprises one of said unique nucleotide sequences, wherein at least one of said nucleotide sequences comprises at least an expression cassette comprising a plant scorable marker gene;
b) co-cultivating said tissue with said Agrobacterium;
c) culturing the tissue in a culture medium comprising an antibiotic capable of inhibiting the growth of Agrobacterium;
d) screening the tissue for expression of said plant scorable marker gene; and
e) regenerating a transformed plant from tissue that expresses said plant scorable marker gene.

Designated contracting States at the time of grant are: Austria, Belgium, Switzerland, Cyprus, Germany, Denmark, Spain, Finland, France, United Kingdom, Greece, Ireland, Italy, Liechtenstein, Luxembourg, Monaco, Netherlands, Portugal, Sweden.
AU 764100

  • Earliest priority – 1 October 1998
  • Filed – 28 September 1999
  • Granted – 7 August 2003
  • Expected expiry – 27 September 2019
 Title – Method of plant transformation

Claim 1
An Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region comprising a heterologous nucleotide sequence of interest.
Claim 12
A method for co-transformation of a plant with at least two unique heterologous nucleotide sequences of interest, said method comprising the steps of:a) contacting a tissue from said plant with an Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region, wherein each of said T-DNA regions comprises one of said unique nucleotide sequences, wherein at least one of said nucleotide sequences comprises at least an expression cassette comprising a plant scorable marker gene;
b) co-cultivating said tissue with said Agrobacterium;
c) culturing the tissue in a culture medium comprising an antibiotic capable of inhibiting the growth of Agrobacterium;
d) screening the tissue for expression of said plant scorable marker gene; and
e) regenerating a transformed plant from tissue that expresses said plant scorable marker gene.
CA 2344700

  • Earliest priority – 1 October 1998
  • Filed – 28 September 1999
  • Granted – 29 March 2005
  • Expected expiry – 27 September 2019
 Title – Method of plant transformation

Claim 1
An Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region comprising a heterologous nucleotide sequence of interest.
Claim 12
A methid for co-transformation of a plant with at least two unique heterologous nucleotide sequences of interest, said method comprising the steps of:a) contacting a tissue from said plant with an Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids, wherein each of said binary vector plasmids comprises at least one T-DNA region, wherein each of said T-DNA regions comprises one of said unique nucleotide sequences, wherein at least one of said nucleotide sequences comprises at least an expression cassette comprising a plant scorable marker gene;
b) co-cultivating said tissue with said Agrobacterium;
c) culturing the tissue in a culture medium comprising an antibiotic capable of inhibiting the growth of Agrobacterium;
d) screening the tissue for expression of said plants scorable marker gene; and
e) regenerating a transformed plant from tissue that expresses said plant scorable marker gene.
WO 2000/18939

  • Earliest priority – 1 October 1998
  • Filed – 28 September 1999
  • OPI – 14 September 2000
 Title – Method of plant transformation

Claim 1
An Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids that are stably inherited, wherein each of said binary vector plasmids comprises at least one T-DNA region comprising a heterologous nucleotide sequence of interest.
Claim 12
A method for co-transformation of a plant with at least two unique heterologous nucleotide sequences of interest, said method comprising the steps of:a) contacting a tissue from said plant with an Agrobacterium strain comprising a helper plasmid and at least two binary vector plasmids that are stably inherited, wherein each of said binary vector plasmids comprises at least one T-DNA region, wherein each of said T-DNA regions comprises one of said unique nucleotide sequences, wherein at least one of said nucleotide sequences comprises at least an expression cassette comprising a plant scorable marker gene;
b) co-cultivating said tissue with said Agrobacterium;
c) culturing the tissue in a culture medium comprising an antibiotic capable of inhibiting the growth of Agrobacterium;
d) screening the tissue for expression of said plants scorable marker gene; and
e) regenerating a transformed plant from tissue that expresses said plant scorable marker gene.

Note: Patent information on this page was last updated on 27 February 2006.