BETA 1,2-xylosyltransferase-gene from arabidopsis
OwnerJosef Glossl, Richard Strasser +5inventors
InventorsJosef Glossl, Richard Strasser, Jan Mucha, Lukas Mach, Friedrich Altmann, Iain Wilson, Herta Steinkellner
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Abstract
A DNA molecule is provided which comprises a sequence according to SEQ ID NO: 8 having an open reading frame from base pair 227 to base pair 1831 or having at least 50% homology to the above-indicated sequence, or hybridizing with the above-indicated sequence under stringent conditions, or comprising a sequence which has degenerated to the above-indicated DNA sequence because of the genetic code, the sequence coding for a plant protein having β1,2-xylosyltransferase activity or being complementary thereto.
Claims46
1-32. (canceled)
An isolated DNA molecule comprising:
- (a) SEQ ID NO:
- 8,
- (b) a sequence that is at least 50% homologous with SEQ ID NO:
- 8,
- (c) a sequence that hybridizes with a complement of SEQ ID NO:
- 8 under stringent conditions,
- (d) a sequence which is degenerate to SEQ ID NO:
- 8 due to the genetic code, or
- (e) a full complement of any one of sequences (a) through (d),
- wherein the sequences (a) through (d) code for a protein having β1,2-xylosyltransferase activity.
A method of preparing a recombinant cell or plant, comprising inserting a mutated β1,2-xylosyltransferase DNA molecule into the genome of said cell or plant by homologous recombination at the position of a non-mutated, β1,2-xylosyltransferase sequence.
A method for producing a recombinant glycoprotein having reduced β1,2-bound xylose residues comprising:
- transfecting a plant cell with a polynucleotide sequence comprising at least 50 nucleotides complementary to a sequence coding for an endogenous protein having β1,2-xylosyltransferase activity, the sequence coding for the endogenous protein having β1,2-xylosyltransferase activity being selected from the group consisting of SEQ ID NO:
- 8, a sequence that is at least 50% identical to SEQ ID NO:
- 8, a sequence that hybridizes to SEQ ID NO:
- 8 under stringent conditions, and a sequence that is degenerate to SEQ ID NO:
- 8 due to the genetic code;
- and
- transfecting the plant cell with a sequence coding for a glycoprotein so that a recombinant glycoprotein having less than 50% of β1,2-bound xylose residues as compared to a recombinant glycoprotein in a non-β1,2-xylosyltransferase reduced plant cell is expressed.
A method for producing a recombinant glycoprotein having reduced β1,2-bound xylose residues comprising:
- obtaining a plant or a plant cell having a deletion, insertion, or substitution in a sequence coding for a protein having β1,2-xylosyltransferase activity, the sequence being selected from the group consisting of SEQ ID NO:
- 8, a sequence that is at least 50% identical to SEQ ID NO:
- 8, a sequence that hybridizes to SEQ ID NO:
- 8 under stringent conditions, and a sequence that is degenerate to SEQ ID NO:
- 8 due to the genetic code, wherein the plant or plant cell has reduced β1,2-xylosyltransferase activity as compared to a plant or plant cell that does not have the deletion, insertion, or substitution in the sequence coding for the protein having β1,2-xylosyltransferase activity;
- and
- transfecting the plant or plant cell with a sequence coding for a glycoprotein so that a recombinant glycoprotein having less than 50% of β1,2-bound xylose residues as compared to a recombinant glycoprotein in a non-β1,2-xylosyltransferase reduced plant or plant cell is expressed.
Dependencies are inferred from the claim text.