Site-specific recombination using an epitope tagged bacteriophage P1 Cre recombinase.
Since the original description of Cre mediated site-specific recombination in bacteriophage P1 (Sternberg, N., Hamilton, D., 1981 J. Mol. Biol., 150, 467-487), the Cre-lox system of recombination has been widely used to manipulate prokaryotic and eukaryotic genomes. Unfortunately, there are few means available to measure Cre protein expression in vivo. We have constructed an expression vector wherein the Cre protein is tagged at the carboxy terminus with an 11-amino-acid epitope to the herpes simplex virus (HSV) glycoprotein D coat protein (Isola, V.J., Eisenberg, R.J., Siebert, G.R., Heilman, C.J., Wilcox, W.C., Cohan, G.H., 1989. J. Virol. 63, 2325-2334). The epitope tag facilitates detection of Cre expression in vitro and in vivo using immunofluorescent labeling with a commercially available antibody. The epitope tag does not interfere with Cre recombinase activity or alter recombination efficiency between loxP sites. We have shown in mice that a transgene expressing our tagged Cre is capable of excising a loxP flanked sequence contributed by another transgenic mouse. In summary, we have developed an epitope-tagged Cre recombinase that is fully active and readily detectable.
Veterans Affairs Medical Center, 500 Foothill Drive, Salt Lake City, UT 84148, USA.
P K
PK
Stricklett
R D
RD
Nelson
D E
DE
Kohan
D016475:3T3 Cells; D000818:Animals; D017101:Bacteriophage P1 / Q000201:enzymology / Q000235:genetics*; D016466:CHO Cells; D006224:Cricetinae; D000939:Epitopes; D019426:Integrases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology*; D051379:Mice; D008822:Mice, Transgenic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D011995:Recombination, Genetic; D018139:Simplexvirus; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis; D014764:Viral Proteins
D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases