Establishment of Double-gene Transgenic Mouse Model for hHT and hDAF through SMGT
Yongfu Chen
Abstract
Yongfu Chen
Abstract
To explore the molecular mechanism of hyperacute rejection(HAR) and delayed xenograft rejection(DXR),double-gene transgenic mouse model expressing Human alpha1,2-fucosyltransferase(hHT) and human-decay accelerating factor(hDAF) was established using sperm-mediated gene transfer optimized by dendrimers(SMGT).The positive rate of PCR for GFP + RFP in 2-cell embryos was 4.51% after the SMGT was optimized,which was significantly higher than that of the control group(P 0.01).Embryos expressing both GFP and RFP were got were transplanted to the pseudopregnant female mice.36 offspring were got.The founder transgenic mice(G0 mice) were identified with PCR and southern blot.The results of PCR showed that genomes from 17 mice were integrated by both hHT and hDAF.The results of southern blot for 5 mice showed a positive signal of hHT + hDAF.These results indicated that G0 double-gene transgenic mouse model for hHT and hDAF was established.
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To explore the molecular mechanism of hyperacute rejection(HAR) and delayed xenograft rejection(DXR),double-gene transgenic mouse model expressing Human alpha1,2-fucosyltransferase(hHT) and human-decay accelerating factor(hDAF) was established using sperm-mediated gene transfer optimized by dendrimers(SMGT).The positive rate of PCR for GFP + RFP in 2-cell embryos was 4.51% after the SMGT was optimized,which was significantly higher than that of the control group(P 0.01).Embryos expressing both GFP and RFP were got were transplanted to the pseudopregnant female mice.36 offspring were got.The founder transgenic mice(G0 mice) were identified with PCR and southern blot.The results of PCR showed that genomes from 17 mice were integrated by both hHT and hDAF.The results of southern blot for 5 mice showed a positive signal of hHT + hDAF.These results indicated that G0 double-gene transgenic mouse model for hHT and hDAF was established.
Key concepts: Transgene, Southern blot, Molecular biology, Biology, Gene, Western blot, Genetically modified mouse, Genetics