Fucosyl transferase gene
AssigneesFriedrich Altmann, Jan Mucha, Haralt Leiter, Josef Glössl, Erika Staudacher
InventorsFriedrich ALTMANN, Jan Mucha, Haralt Leiter, Josef Glossl, Erika Staudacher
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Abstract
A DNA molecule is provided which comprises a sequence according to SEQ ID NO: 1 having an open reading frame from base pair 211 to base pair 1740 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 fucosyltransferase activity or being complementary thereto.
Claims15
-42. (canceled)
A method of producing a recombinant glycoprotein, comprising expressing a recombinant glycoprotein in plants or plant cells, wherein an endogenous α1,3-fucosyltransferase production is suppressed or completely stopped,
- wherein said endogenous α1,3-fucosyltransferase is identified by sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO:
- 1 with an open reading frame from base pair 211 to base pair 1740, and at least suppressing said endogenous α1,3-fucosyltransferase production.
The method of claim 1, wherein said endogenous α1,3-fucosyltransferase can be identified by sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO:
- 1 with an open reading frame from base pair 211 to base pair 1740 by the program fastDB.
The method according to claim 1, wherein the glycoprotein is a human protein.
The method of claim 1, wherein the expression of the α1,3-fucosyltransferase is suppressed or completely blocked by a knock-out mutation of the endogenous α1,3-fucosyltransferase gene in said plant or plant cell.
The method of claim 1, wherein the expression of the α-1,3-fucosyltransferase is suppressed or completely blocked by antisense inhibition in said plant or plant cell.
The method of claim 1, wherein the expression of the α1,3-fucosyltransferase is suppressed or completely blocked by transfection with a polynucleotide comprising a sequence of at least 50 nucleotides which is complementary to the sequence coding for the endogenous α1,3-fucosyltransferase.
The method of claim 1, wherein the endogenous α1,3-fucosyltransferase activity in said plant or plant cell is less than 50% of the α1,3-fucosyltransferase occurring in natural plants or plant cells without α1,3-fucosyltransferase suppression.
A method of producing a recombinant glycoprotein, comprising expressing a recombinant glycoprotein in plants or plant cells, wherein an endogenous α1,3-fucosyltransferase production is suppressed or completely stopped, comprising identifying an endogenous α1,3-fucosyltransferase by (i) sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO:
- 1 with an open reading frame from base pair 211 to base pair 1740 and (ii) assay for α1,3-fucosyltransferase activity, and suppressing or completely stopping said endogenous α1,3-fucosyltransferase production.
A method of producing plants or plant cells with reduced or stopped endogenous α1,3-fucosyltransferase activity, comprising identifying an endogenous α1,3-fucosyltransferase by (i) sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO:
- 1 with an open reading frame from base pair 211 to base pair 1740 and (ii) assay for α1,3-fucosyltransferase activity, and suppressing or completely stopping said endogenous α1,3-fucosyltransferase production.
The method of claim 9, wherein said endogenous α1,3-fucosyltransferase can be identified by sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO:
- 1 with an open reading frame from base pair 211 to base pair 1740 by the program fastDB.
The method of claim 9, wherein the expression of the α1,3-fucosyltransferase is suppressed or completely blocked by a knock-out mutation of the endogenous α1,3-fucosyltransferase gene in said plant or plant cell.
The method of claim 9, wherein the expression of the α1,3-fucosyltransferase is suppressed or completely blocked by antisense inhibition in said plant or plant cell.
The method of claim 9, wherein the expression of the α1,3-fucosyltransferase is suppressed or completely blocked by transfection with a polynucleotide comprising a sequence of at least 50 nucleotides which is complementary to the sequence coding for the endogenous α1,3-fucosyltransferase.
The method of claim 9, wherein the endogenous α1,3-fucosyltransferase activity in said plant or plant cell is less than 50% of the α1,3-fucosyltransferase occurring in natural plants or plant cells without α1,3-fucosyltransferase suppression.
Dependencies are inferred from the claim text.