DNA encoding human cytoplasmic Cu/Zn superoxide dismutase
AssigneeChiron Corporation
InventorsRobert A. Hallewell, Guy T. Mullenbach
Life of this patent
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Abstract
Methods and compositions are provided for the production of human superoxide dismutase and a novel protocol for enhancing efficiency of expression. The gene encoding for human superoxide dismutase is isolated and inserted into a vector in conjunction with a synthetic linker which provides for enhanced efficiency in translation. E. coli strain D1210 (pSODX8) was deposited at the A.T.C.C. on Sep. 27, 1983 and given Accession No. 39453. Yeast strain 2150-2-3 (pC1/1GAPSOD) and E. coli strains D1210 (pSOD11) and D1210 (pS2OR) were deposited at the A.T.C.C. on May 9, 1984, and given Accession Nos. 20708, 39679 and 39,680, respectively.
Claims28
An intron-free DNA molecule which comprises a coding sequence for human cytoplasmic Cu/Zn superoxide dismutase, said intron-free DNA molecule comprising a synthetic linker sequence that (a) encodes at least amino acids 1-9 of the amino acid sequence of human Cu/Zn superoxide dismutase and (b) comprises a DNA spacer sequence in a region between the ribosomal binding site and the first codon for said superoxide dismutase coding sequence, wherein said linker sequence is in reading frame with a cDNA fragment that encodes the remainder of the amino acid sequence of human cytoplasmic Cu/Zn superoxide dismutase, wherein the coding sequence of the DNA molecule comprises the following nucleotide sequence:
- 5'-ATG GCX ACX AAG GCT GTT TGT GTT TTG AAG GGT GAC GGC CCA GTT CAA GGT ATT ATT AAC TTC GAG CAG AAG GAA AGT AAT GGA CCA GTG AAG GTG TGG GGA AGC ATT AAA GGA CTG ACT GAA GGC CTG CAT GGA TTC CAT GTT CAT GAG TTT GGA GAT AAT ACA GCA GGC TGT ACC AGT GCA GGT CCT CAC TTT AAT CCT CTA TCC AGA AAA CAC GGT GGG CCA AAG GAT GAA GAG AGG CAT GTT GGA GAC TTG GGC AAT GTG ACT GCT GAC AAA GAT GGT GTG GCC GAT GTG TCT ATT GAA GAT TCT GTG ATC TCA CTC TCA GGA GAC CAT TGC ATC ATT GGC CGC ACA CTG GTG GTC CAT GAA AAA GCA GAT GAC TTG GGC AAA GGT GGAAAT GAA GAAAGT ACA AAG ACA GGA AAC GCT GGC AGT CGT TTG GCT TGT GGT GTA ATT GGG ATC GCC CAA TAA-3';
- wherein GCX represents GCA or GCT;
- and ACX represents ACG.
An intron-free double-stranded DNA molecule wherein one strand of the DNA molecule comprises a coding sequence for human cytoplasmic Cu/Zn superoxide dismutase, said one strand comprising a synthetic linker sequence that (a) encodes at least amino acids 1-9 of the amino acid sequence of human Cu/Zn superoxide dismutase and (b) comprises a DNA spacer sequence in a region between the ribosomal binding site and the first codon for said superoxide dismutase coding sequence, wherein said linker sequence is in reading frame with a cDNA fragment that encodes the remainder of the amino acid sequence of human cytoplasmic Cu/Zn superoxide dismutase, wherein the coding sequence of the DNA molecule comprises the following nucleotide sequence:
- 5'-ATG GCX ACX AAG GCT GTT TGT GTT TTG AAG GGT GAC GGC CCA GTT CAA GGT ATT ATT AAC TTC GAG CAG AAG GAA AGT AAT GGA CCA GTG AAG GTG TGG GGA AGC ATT AAA GGA CTG ACT GAA GGC CTG CAT GGA TTC CAT GTT CAT GAG TTT GGA GAT AAT ACA GCA GGC TGT ACC AGT GCA GGT CCT CAC TTTAAT CCT CTA TCC AGA AAA CAC GGT GGG CCA AAG GAT GAA GAG AGG CAT GTT GGA GAC TTG GGC AAT GTG ACT GCT GAC AAA GAT GGT GTG GCC GAT GTG TCT ATT GAA GAT TCT GTG ATC TCA CTC TCA GGA GAC CAT TGC ATC ATT GGC CGC ACA CTG GTG GTC CAT GAA AAA GCA GAT GAC TTG GGC AAA GGT GGA AAT GAA GAA AGT ACA AAG ACA GGA AAC GCT GGC AGT CGT TTG GCT TGT GGT GTA ATT GGG ATC GCC CAA TAA-3';
- wherein GCX represents GCA or GCT;
- and ACX represents ACG.
Dependencies are inferred from the claim text.