Asselbergs F A, Pronk R
Biotechnology Department, CIBA-GEIGY Ltd., Basle, Switzerland.
Mol Biol Rep. 1992 Nov;17(1):61-70. doi: 10.1007/BF01006400.
Shuttle vectors expressing resistance to hygromycin B in both E. coli and in mammalian cells were constructed. A combination of the simian virus 40 early promoter upstream of the native bacterial promoter of the neo gene from transposon Tn5 was found to express hygromycin B resistance better in both types of host cells than a combination of the Tn5 promoter followed by the promoter of the Herpes simplex virus thymidine kinase gene. Hygromycin phosphotransferase fusion proteins with extensions at the carboxyterminus were also tested and found to be marginally less effective as selection markers in eukaryotic cells but virtually inactive in E. coli.
构建了在大肠杆菌和哺乳动物细胞中均表达对潮霉素B抗性的穿梭载体。发现来自转座子Tn5的neo基因天然细菌启动子上游的猿猴病毒40早期启动子的组合,在两种宿主细胞中比Tn5启动子与单纯疱疹病毒胸苷激酶基因启动子的组合能更好地表达潮霉素B抗性。还测试了在羧基末端带有延伸的潮霉素磷酸转移酶融合蛋白,发现其作为真核细胞中的选择标记效果略差,但在大肠杆菌中几乎无活性。