Breeding and genotype identification of IP3R2 knockout mice
Objective Breeding and identify IP3R2 knockout mice,to provide sample support for the subsequent experiments,and simultaneous validation of PCR applicability for gene identification.Method Fifteen IP3R2 knockout heterozygous mice were purchased and bred in a combined cage of 1 male and 2 females,When the propagated offspring mice were grown to 6 weeks,the shear mouse tail was used for gene extraction and PCR amplification,and the results of agarose gel electrophoresis were determined.Brain samples from homozygous IP3R2 knockout mice and wild-type were indentified for western blot and immunofluorescence to verify the expression of IP3R2 protein in the brain tissue.Daughter mouse brain tissues were harvested for Western blot and immunofluorescence to verify the expression of IP3R2 protein expression in homozygous IP3R2 knockout mice.Results The IP3R2 gene knockout mice can be success-fully bred.In the offspring mice,IP3R2 homozygosity was 23.3%.PCR amplification can successfully identify the genotypes of the prog-eny mice.Compared to the wild mice,IP3R2 expression was significantly lower in IP3R2 knockout mice(P<0.05).Conclusion This experiment successfully established and identified the IP3R2 gene knockout mice,to provide a large number of samples for subsequent experiments,and the PCR technology can quickly and accurately identify the IP3R2 genotype,to provide technical support for the subse-quent experiments.
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