[The study of a HPRT locus-specific transgenic method based on FLP recombinase mediated cassette exchange].
Exchange plasmid pF-HPRT-F3 and Flp expression plasmid pCMV-Flp were constructed and then introduced using electroporation system into F18 ES cell line, which have an exchange cassette F-Neo-F3 at HPRT locus. After HAT selection, HAT resistant clones were obtained. Then G418 sensitivity test and Southern blotting were carried out to screen RMCE recombinants. The results indicated that RMCE had taken place in three of 12 HAT resistant clones. The frequency is 25%. The result demonstrates that it is realizable to introduce transgene to HPRT locus by using Flp recombinase mediated cassette exchange reaction.
Zheng
J M
JM
Department of Medical Genetics, Second Military Medical University, Shanghai 200433, China.
He
Y
Y
Fu
J L
JL
D000818:Animals; D002460:Cell Line; D002999:Clone Cells; D004254:DNA Nucleotidyltransferases / Q000502:physiology*; D018274:Electroporation; D005822:Genetic Vectors; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D051379:Mice; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D014162:Transfection
D007041:Hypoxanthine Phosphoribosyltransferase; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase