A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure.
A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure.
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突变的新霉素磷酸转移酶 II 基因降低了转化体对抗生素选择压力的抗性。
DOI：
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发表时间：
1990
影响因子：
11.1
通讯作者：
John W. Pellow
中科院分区：
文献类型：
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作者：
R. Yenofsky;Miriam Fine;John W. Pellow
转座子Tn5的neo（新霉素耐药）基因编码新霉素磷酸转移酶II (EC 2.7.1.95)，使其对多种氨基糖苷类抗生素产生耐药性，包括卡那霉素和G418。该基因被广泛用作细菌、酵母、植物和动物等多种生物转化的选择性标记。我们在几种常用转化载体的嵌合新霉素磷酸转移酶II基因编码的蛋白中发现了一个突变，涉及在残基182处谷氨酸到天冬氨酸的转化。该突变显著降低了磷酸转移酶的活性，但似乎不影响新霉素磷酸转移酶II mRNA或蛋白的稳定性。用突变基因转化的植物和细菌对抗生素的耐药性低于用正常基因转化的植物和细菌。一个简单的限制性内切酶酶切区分突变基因和正常基因。
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.