BETA 1,2-xylosyltransferase-gene from arabidopsis

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.

1 - 32 . (canceled)  
     
     
         33 . 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.    
     
     
         34 . The isolated DNA molecule according to  claim 33 , wherein said sequence is at least 70% identical to the sequence of SEQ ID NO. 8.  
     
     
         35 . The isolated DNA molecule according to  claim 33 , wherein said sequence is at least 80% identical to the sequence of SEQ ID NO. 8.  
     
     
         36 . The isolated DNA molecule according to  claim 33 , wherein the isolated DNA molecule comprises 1780 to 1880 nucleotides.  
     
     
         37 . The isolated DNA molecule according to  claim 33 , covalently associated with a detectable marker substance.  
     
     
         38 . An isolated DNA molecule coding for a ribozyme, wherein the isolated DNA molecule has two sequence sections, each of which has a length of at least 10 to 15 nucleotides and is complementary to sequence sections of a DNA molecule according to  claim 33  so that said ribozyme complexes and cuts the mRNA transcribed by a natural β1,2-xylosyltransferase DNA molecule.  
     
     
         39 . A biologically functional vector, comprising a DNA molecule according to  claim 33  and a promoter.  
     
     
         40 . A biologically functional vector comprising a DNA molecule according to  claim 33  and a promoter, wherein said DNA molecule is inversely orientated with respect to the promoter.  
     
     
         41 . A method of producing a β1,2-xylosyltransferase, comprising cloning a DNA molecule according to  claim 33  into a vector, transfecting said vector into a host cell, and selecting and amplifying transfected host cells, wherein the host cell expresses the β1,2-xylosyltransferase.  
     
     
         42 . A method of suppressing endogenous β1,2-xylosyltransferase in a plant cell comprising, inserting a vector according to  claim 39  into the plant cell, wherein the production of β1,2-xylosyltransferase is suppressed.  
     
     
         43 . A method of suppressing endogenous β1,2-xylosyltransferase in a plant cell comprising, inserting a vector according to  claim 40  into the plant cell, wherein the production of β1,2-xylosyltransferase is suppressed.  
     
     
         44 . 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.  
     
     
         45 . The method of  claim 44 , wherein β1,2-xylosyltransferase production is suppressed in the recombinant cell or plant.  
     
     
         46 . The method of  claim 44 , wherein the mutation is a deletion, insertion, or substitution mutation.  
     
     
         47 . A PNA molecule comprising a nucleotide sequence complementary to the sequence of a DNA molecule according to  claim 33 .  
     
     
         48 . A PNA molecule comprising a nucleotide sequence corresponding to the sequence of a DNA molecule according to  claim 33 .  
     
     
         49 . A method of producing plants or plant cells having suppressed expression of β1,2-xylosyltransferase at the transcription or translation level, comprising inserting PNA molecules according to  claim 47  into the plants or plant cells.  
     
     
         50 . A method of producing recombinant glycoproteins, comprising transfecting recombinant plants or plant cells according to  claim 45  with a gene that expresses the glycoprotein so that the recombinant glycoproteins are expressed.  
     
     
         51 . The method of  claim 50 , wherein the recombinant glycoproteins are recombinant human glycoproteins.  
     
     
         52 . A method of selecting DNA molecules coding for a β1,2-xylosyltransferase in a sample, comprising adding DNA molecules according to  claim 37  to said sample, which molecules bind to the DNA molecules coding for a β1,2-xylosyltransferase.  
     
     
         53 . The method of  claim 52 , wherein said sample comprises genomic DNA of a plant or non-vertebrate animal organism.  
     
     
         54 . A preparation of β1,2-xylosyltransferase produced by a method according to  claim 41 , wherein said preparation has isoforms having pI values of between 6.0 and 9.0.  
     
     
         55 . The preparation of  claim 54 , wherein said preparation has an isoform having a pI value of 7.52.  
     
     
         56 . A method of preparing a plantified carbohydrate unit of a vertebrate glycoprotein comprising adding to a sample comprising a carbohydrate unit, a glycoconjugate or a glycoprotein, a β1,2-xylosyltransferase encoded by a DNA molecule according to  claim 33  and a UDP-xylose, so that xylose is bound to said carbohydrate unit, glycoconjugate or glycoprotein, at the β1,2-position by said β1,2-xylosyltransferase.  
     
     
         57 . The method of  claim 56 , wherein the vertebrate glycoprotein is a human glycoprotein.  
     
     
         58 . 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.    
     
     
         59 . The method of  claim 58 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 70% identical to SEQ ID NO: 8.  
     
     
         60 . The method of  claim 58 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 80% identical to SEQ ID NO: 8.  
     
     
         61 . The method of  claim 58 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 95% identical to SEQ ID NO: 8.  
     
     
         62 . The method of  claim 58 , wherein the recombinant glycoprotein has less than 20% of β1,2-bound xylose residues as compared to a recombinant glycoprotein in a non-xylosyltransferase reduced plant cell.  
     
     
         63 . The method of  claim 58 , wherein the recombinant glycoprotein has no β1,2-bound xylose residues.  
     
     
         64 . The method of  claim 58 , wherein the polynucleotide sequence is an antisense RNA.  
     
     
         65 . The method of  claim 58 , wherein the polynucleotide sequence is a double-stranded RNA.  
     
     
         66 . The method of  claim 58 , wherein the polynucleotide sequence comprises a PNA.  
     
     
         67 . The method of  claim 58 , wherein the recombinant glycoprotein is a human glycoprotein.  
     
     
         68 . 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.    
     
     
         69 . The method of  claim 68 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 70% identical to SEQ ID NO: 8.  
     
     
         70 . The method of  claim 68 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 80% identical to SEQ ID NO: 8.  
     
     
         71 . The method of  claim 68 , wherein the sequence that is at least 50% identical to SEQ ID NO: 8 is a sequence that is at least 95% identical to SEQ ID NO: 8.  
     
     
         72 . The method of  claim 68 , wherein the recombinant glycoprotein has less than 20% of β1,2-bound xylose residues as compared to a recombinant glycoprotein in a non-xylosyltransferase reduced plant cell.  
     
     
         73 . The method of  claim 68 , wherein the recombinant glycoprotein has no β1,2-bound xylose residues.  
     
     
         74 . The method of  claim 68 , wherein the polynucleotide sequence is an antisense RNA.  
     
     
         75 . The method of  claim 68 , wherein the polynucleotide sequence is a double-stranded RNA.  
     
     
         76 . The method of  claim 68 , wherein the polynucleotide sequence comprises a PNA.  
     
     
         77 . The method of  claim 68 , wherein the recombinant glycoprotein is a human glycoprotein.