Patent PL-205710-B1 (PL)
Title: Recombined glycoproteins, DNA molecules, biologically active vector, method for cDNA obtaining, method for GlcNAc 1, 3-fucosyltransferase cloning, method for obtaining recombined host cells, method for obtaining recombined plants, recombined plants and plant cells, recombined insects and insect cells, peptide nucleid acid molecules, method for obtaining plants or insects of cells thereof with blocked expression of GlcNAc 1, 3-fucosyltransferase, method for obtaining recombined glycoproteins and human glycoproteins, reco
Inventor: ALTMANN FRIDRICH
IPC: C12N15/11
Filed: 2000-02-17
Abstract: A DNA molecule (I), encoding a plant protein with fucosyl transferase activity, comprising a 2198 base pair sequence, fully defined in the specification, with an open reading frame of nucleotides 211-1740, a sequence at least 50 % homologous to it, a sequence hybridizing to it under stringent conditions, a sequence which differs due to genetic code degeneracy, or their complement, is new. Independent claims are also included for the following: (1) a DNA molecule (II), comprising a 105 base pair sequence, fully defined in the specification, a sequence at least 95 % homologous to it, a sequence that hybridizes to it under stringent conditions, or a sequence which differs due to genetic code degeneracy; (2) a DNA molecule which is a partial sequence of (I) or (II), which comprises 30-50 base pairs; (3) (I) or (II) covalently associated with a detectable marker substance; (4) a biologically functional vector comprising (I) or (II) or a partial sequence of at least 20 base pairs; (5) a biologically functional vector comprising (I), (II) or a partial sequence in inverse orientation to the promoter; (6) (I), (II) or the DNA molecule of (2) or (3) comprising a deletion, insertion and/or substitution mutation; (7) a DNA encoding a ribozyme, which comprises two sequence sections of at least 10-15 base pairs, which are complementary to (I) or (II), so that the ribozyme complexes with and cuts the mRNA, which is transcribed from a natural GlcNAc-alpha 1,3-fucosyl transferase DNA molecule; (8) a biologically functional vector comprising DNA of (6) or (7); (9) producing a cDNA of (I) or (II) comprising isolating RNA of insects or plants, and carrying out reverse transcription with a reverse transcriptase and primers; (10) cloning a GlcNAc-alpha 1,3-fucosyl transferase; (11) producing recombinant host cells which over-express GlcNAc-alpha 1,3-fucosyltransferase; (12) producing recombinant host cells, preferably plant or insect cells, which have a DNA of (6) in place of the non-mutated homologous sequence; (13) a recombinant plant or insect cell produced by the method of (12) and which over-expresses the GlcNAc-alpha 1,3-fucosyltransferase; (14) a PNA (peptide-nucleic acid) molecule which is complementary to or the same as (I); (15) producing a plant or insect cell with blocked GlcNAc-alpha 1,3-fucosyl transferase expression at the level of transcription or translation by the PNA of (14); (16) a method of recombinant glycoprotein production; (17) a recombinant glycoprotein produced in a plant or insect cell system as above; a GlcNAc-alpha 1,3-fucosyltransferase preparation, cloned by a method of (16), where the isoforms have a pI of between 6 and 9, preferably 6.8 and 8.2; and (18) producing human and other vertebrate carbohydrates by transplantation.