RRC ID 30935
Author Niwa H, Yamamura K, Miyazaki J.
Title Efficient selection for high-expression transfectants with a novel eukaryotic vector.
Journal Gene
Abstract We have developed a new expression vector which allows efficient selection for transfectants that express foreign genes at high levels. The vector is composed of a ubiquitously strong promoter based on the beta-actin promoter, a 69% subregion of the bovine papilloma virus genome, and a mutant neomycin phosphotransferase II-encoding gene driven by a weak promoter, which confers only marginal resistance to G418. Thus, high concentrations of G418 (approx. 800 micrograms/ml) effectively select for transfectants containing a high vector copy number (greater than 300). We tested this system by producing human interleukin-2 (IL-2) in L cells and Chinese hamster ovary (CHO) cells, and the results showed that high concentrations of G418 efficiently yielded L cell and CHO cell transfectants stably producing IL-2 at levels comparable with those previously attained using gene amplification. The vector sequences were found to have integrated into the host chromosome, and were stably maintained in the transfectants for several months.
Volume 108(2)
Pages 193-9
Published 1991-12-15
DOI 10.1016/0378-1119(91)90434-d
PII 0378-1119(91)90434-D
PMID 1660837
MeSH Actins / genetics Animals Blotting, Northern Blotting, Southern Bovine papillomavirus 1 / genetics* CHO Cells Cattle Cricetinae Genetic Vectors / genetics* Gentamicins / pharmacology Humans Interleukin-2 / genetics Kanamycin Kinase L Cells Mice Phosphotransferases / genetics Promoter Regions, Genetic / genetics Recombinant Proteins / biosynthesis* Recombinant Proteins / genetics Selection, Genetic Transfection / genetics beta-Galactosidase / genetics beta-Galactosidase / metabolism
IF 2.984
Times Cited 4324
DNA material pCAGGS (RDB08938)