Construction of a neo fusion gene for expression in both prokaryotic and eukaryotic cells.
A high-copy-number plasmid, pLink, was constructed to allow the direct selection in Escherichia coli of a neo fusion gene capable of conferring Geneticin (G418) resistance on mouse L cells. pLink was derived from pdMmtneo by insertion of a KpnI linker within the 5'-coding region of the neo gene. This created a minus-one frameshift mutation resulting in a translational termination within the N-terminal region of the protein. The Neo activity was restored by insertion into the modified neo gene of a piece of coding sequence derived from human HPRT cDNA. The resulting plasmid, pAH, was microinjected into mouse A9 cells and shown to confer resistance to G418.
Harwood
A J
AJ
AFRC Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, U.K.
Shervington
A
A
Bostock
C J
CJ
D000818:Animals; D001483:Base Sequence; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005839:Gentamicins; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009355:Neomycin; D010443:Peptide Chain Termination, Translational / Q000187:drug effects; D010957:Plasmids; D011993:Recombinant Fusion Proteins; D012641:Selection, Genetic
D005839:Gentamicins; D011993:Recombinant Fusion Proteins; C010680:antibiotic G 418; D007041:Hypoxanthine Phosphoribosyltransferase; D009355:Neomycin