Gao Chuan, Zhu Xu-Dong, Zhou Xiao-Wei, Yu Fang, Lu Bai-Song, Huang Pei-Tang
Institute of Pharmaceutical Chemistry, Beijing 102205, China.
Sheng Wu Gong Cheng Xue Bao. 2002 May;18(3):308-12.
Neomycin-resistance gene is widely used as a selectable marker in eukaryotic expression vector. It codes neomycin phosphotransferase II (NPT II) which confers resistance to various aminoglycoside antibiotic such as G418 and kanamycine. In this work, by site-directed mutagenesis the neo gene mutant was obtained. The expression vector pmDNA using the neo gene mutant as selectable marker has been constructed. After inserting interest luciferase gene, the expression plasmid pmDNAluc + was stably transfected CHO-K1 cells. As a result, the expression positive ratio reaches to approximate 95% and the ratio of high expression colonies is apparently higher than the controls.
新霉素抗性基因在真核表达载体中被广泛用作选择标记。它编码新霉素磷酸转移酶II（NPT II），赋予对各种氨基糖苷类抗生素如G418和卡那霉素的抗性。在这项工作中，通过定点诱变获得了neo基因突变体。构建了以neo基因突变体为选择标记的表达载体pmDNA。插入目的荧光素酶基因后，表达质粒pmDNAluc +被稳定转染到CHO-K1细胞中。结果，表达阳性率达到约95%，高表达菌落的比例明显高于对照。
