[Construction of eukaryotic expression vector using neomycin-resistance gene mutant as selectable marker].
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.
Gao
Chuan
C
Institute of Pharmaceutical Chemistry, Beijing 102205, China. g.ch.chuan@263.net
Zhu
Xu-Dong
XD
Zhou
Xiao-Wei
XW
Yu
Fang
F
Lu
Bai-Song
BS
Huang
Pei-Tang
PT
D000595:Amino Acid Sequence; D001483:Base Sequence; D004351:Drug Resistance / Q000235:genetics*; D005819:Genetic Markers; D005822:Genetic Vectors; D019868:Kanamycin Kinase / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation
D005819:Genetic Markers; D019868:Kanamycin Kinase