Universal dominant selectable marker cassette
AssigneeCetus Corporation
InventorsDavid H. Gelfand, Frances C. Lawyer, Susanne Stoffel
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Abstract
A novel universal dominant selectable marker cassette is disclosed. The marker comprises the coding sequences for aminoglycoside phosphotransferase I (APH-I) which has been modified and truncated so as to render its use in recombinant vectors more convenient. The modified, truncated sequence (mtAPH-I) gene is capable, upon expression, of conferring resistance to a number of antibiotics on the host. One of these antibiotics, G418, is toxic to eucaryotic as well as procaryotic hosts. Also disclosed are methods of constructing fusion proteins having N-terminal sequences corresponding to a desired peptide sequence, and C-terminal sequences comprising the amino acids encoded by mtAPH-I.
Claims29
A DNA sequence cassette encoding a dominant selectable marker effective in procaryotic and in eucaryotic transformants which comprises:
- 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 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 duplicative of said cassette-unique restriction site, and truncated by deletion of codons encoding at least one amino acid in positions 2-10 inclusive.
Dependencies are inferred from the claim text.