[Detection of Cre recombinase activity in Mx-Cre transgenic mice induced by INF in vitro].
The Cre recombinase and its activity in C57-TgN(Mx-Cre) transgenic mice is studied by polymerase chain reaction (PCR), Western blot, immunohistochemistry, immunogold electron microscopy and Southern blot. C57-TgN(Mx-Cre) transgenic mice harbouring cre gene in genomic DNA is demonstrated by PCR, and these mice which are induced by INF-alpha 1b could express Cre recombinase, which is confirmed by Western blot. With immunohistochemistry, we find that the Cre recombinase expresses in hepatocyte cytoplasm and nuclear of C57-TgN(Mx-Cre) transgenic mice. Cre recombinase expressed in hepatocyte cytoplasm and nuclear is further confirmed by immunogold electon microscopy. And it is supported that the Cre recombinase which is created from C57-TgN(Mx-Cre) transgenic mice induced by INF-alpha 1b can direct DNA recombination reaction in vitro. All evidence leads us supporting the view that the Cre recombinase expressed in C57-TgN(Mx-Cre) transgenic mice has activity. Thus we find a method to detect the activity of Cre recombinase in vitro.
Zi
X Y
XY
Department of Cell Biology, Second Military Medical University, Shanghai 200433, China.
Yao
Y C
YC
Xiong
J
J
Li
J X
JX
Yan
Y B
YB
Yu
H Y
HY
Hu
Y P
YP
D000818:Animals; D004790:Enzyme Induction; D019204:GTP-Binding Proteins; D019426:Integrases / Q000378:metabolism*; D007372:Interferons / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D064547:Myxovirus Resistance Proteins; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000502:physiology*; D014764:Viral Proteins / Q000378:metabolism*
C579070:Mx1 protein, mouse; D064547:Myxovirus Resistance Proteins; D011506:Proteins; D014764:Viral Proteins; D007372:Interferons; C045073:Cre recombinase; D019426:Integrases; D019204:GTP-Binding Proteins