Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1988-07-22
1992-05-26
Robinson, Douglas W.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
435232, 435190, 4351723, 435 697, 530350, C12N 910, C12N 904, C12N 1500
Patent
active
051167506
ABSTRACT:
A novel a selectable fusion protein having aminoglycoside phosphotransferase activity 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. The preferred N-terminal sequences are the first 11 amino acids of .beta.-isopropyl malate dehydrogenase, and the first 7 amino acids of yeast enolase.
REFERENCES:
Southern, P. J. et al., "Transformation of Mammalion Cells to Antibiotic Resistance with a Bacterial Gene . . . " J. Mol. Appl. Genetics, vol. 1, pp. 327-341, 1982.
Martinez-Arias, A. E. et al., "Fusion of the Saccharomyces Cerevisiae leu 2 Gene to an Escherichia coli B-Galactosidase Gene," mol. Cell. Biology, vol. 3, No. 4, pp. 580-586, 1983.
Colbene-Garapin, F. et al., "A New Dominant Hybrid Selective Marker for Higher Eukaryotic Cells", J. Mol. Biol., 150, pp. 1-14, 1981.
Jimenez, A et al., "Expression of a Transposable Antibiotic Resistance Element in Saccharomyces", Nature, vol. 267, pp. 869-871, 1980.
Gelfand David H.
Lawyer Frances C.
Stoffel Susanne
Cetus Corporation
Giotta Gregory J.
Halluin Albert P.
Knode Marian C.
Robinson Douglas W.
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