Patents granted to Cetus Corporation (US patent) and Chiron Corporation (CA patent) – Actual granted independent claims

Patents granted to Cetus Corporation (U.S. patent) & Chiron Corporation (CA patent), Actual granted independent claims

US 4784949* & CA 1337716 A1
(* Only Claim 1Claim 1
A DNA sequence cassette encoding a dominant selectable marker effective in prokaryotic and eukaryotic transformants which comprises:

a) an ATG start codon in reading frame with the codons of a modified truncated aminoglycoside phosphotransferase-I (aph-I) gene, designated herein mtaph-I, and
b) at least one cassette-unique restriction site upstream of and proximal to the ATG start codon, said mtaph-I defined herein as the coding sequence for aph -I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of said cassette-unique restriction site, and
truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

Claim 4A recombinant expression vector capable of conferring resistance to G418 on a prokaryotic or eukaryotic transformant which vector comprises:

  • an ATG start codon in reading frame with the codons of the mtaph-I gene, all operably linked to appropriate control sequences functional in the host harboring the vector,
    said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and
    truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.
Claim 17An expression vector, operable in eukaryotic host cells, which comprises:

  • mtaph-I coding sequences operably linked to control sequences functional in the eucaryotic host,

    said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and
    truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

Claim 19An expression vector, operable in eucaryotic host cells, which comprises:

  • a coding sequence for a fusion protein operably linked to control sequences functional in the eucaryotic host,
    wherein the encoded fusion protein comprises:
    a) a desired peptide as the N-terminal sequence and
    b) mtaph-I as the C-terminal sequence, said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and
    truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.
Claim 30A fusion protein which comprises:

a) an N-terminal sequence comprising the N-terminal amino acid sequence of a desired protein and
b) a C-terminal sequence comprising mtaph-I, said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and
truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

Claim 31A fusion protein which comprises:

a) an N-terminal sequence comprising the N-terminal amino acid sequence of B-isopropylmalate dehydrogenase and
b) a C-terminal sequence comprising mtaph-I, said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and
truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

Claim 35A fusion protein which comprises:

a) an N-terminal sequence comprising the N-terminal amino acid sequence of yeast enolase and
b) a C-terminal sequence comprising mtaph-I, said mtaph-I defined herein as the coding sequence for aph-I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of a cassette-unique restriction site, and truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

Claim 37A method of purifying a fusion protein which method comprises:

  • subjecting a fusion protein containing mtaph-I to an amino glycoside containing support, said mtaph-I defined herein as the coding sequence for aph -I modified so as to retain the amino acid sequence encoded by the aph-I gene, but to eliminate any restriction site otherwise duplicated of said cassette-unique restriction site, and truncated by deletion of codons encoding at least one amino acid in positions 2 – 10 inclusive.

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