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Publication : A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure.

First Author  Yenofsky RL Year  1990
Journal  Proc Natl Acad Sci U S A Volume  87
Issue  9 Pages  3435-9
PubMed ID  2159150 Mgi Jnum  J:91743
Mgi Id  MGI:3050526 Doi  10.1073/pnas.87.9.3435
Citation  Yenofsky RL, et al. (1990) A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc Natl Acad Sci U S A 87(9):3435-9
abstractText  The neo (neomycin-resistance) gene of transposon Tn5 encodes the enzyme neomycin phosphotransferase II (EC 2.7.1.95), which confers resistance to various aminoglycoside antibiotics, including kanamycin and G418. The gene is widely used as a selectable marker in the transformation of organisms as diverse as bacteria, yeast, plants, and animals. We found a mutation that involves a glutamic to aspartic acid conversion at residue 182 in the protein encoded by the chimeric neomycin phosphotransferase II genes of several commonly used transformation vectors. The mutation substantially reduces phosphotransferase activity but does not appear to affect the stability of the neomycin phosphotransferase II mRNA or protein. Plants and bacteria transformed with the mutant gene are less resistant to antibiotics than those transformed with the normal gene. A simple restriction endonuclease digestion distinguishes between the mutant and the normal gene.
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