Epitope tagging genomic DNA using a CD-tagging Tn10 minitransposon.
Here, we describe an efficient system for epitope tagging cloned genes by CD tagging using a mini-Tn10 transposon delivery vector. The system was tested against a lambdaFIX genomic clone of the human nucleolin gene. Transfection of HeLa cells with the tagged gene led to the expression of both the appropriately spliced tagged transcript and the appropriately localized tagged protein.
Telmer
C A
CA
Carnegie Mellon University, Pittsburgh, PA, USA.
Berget
P B
PB
Ballou
B
B
Murphy
R F
RF
Jarvik
J W
JW
D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000032:analysis* / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008164:Luminescent Proteins / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D019895:Transposases / Q000235:genetics; D000097796:Nucleolin
D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D019895:Transposases; D000097796:Nucleolin