动物造模,基因敲除 z-bio 2021-12-13 283

　　Objective: To explore the identification method and optimal breeding method of caveolin-1 gene knockout mice, and to provide an ideal animal model for in-depth study of the role of caveolin-1 in the repair of cerebral ischemia injury.

　　Methods: The introduced caveolin-1 gene knockout mice were raised in SPF-level laboratories, and the genomic DNA of rat tail tissues was extracted by boiling and lysis method, and the genotype was detected by PCR reaction based on the primer sequence provided by the Jackson Laboratory in the United States , Using caveolin-1+/- heterozygous crosses, heterozygous and caveolin-1-/- homozygous crosses (orthogonal and reverse crosses), and homozygous crosses 4 different mating methods to observe the conception of parental mice Rate, appearance characteristics and homozygous rate of offspring mice.

　　Results: Agarose gel electrophoresis showed that the molecular weights of PCR products were about 200 bp and 661 bp, which were consistent with the expected molecular weights of the target gene fragments. Different genotypes of caveolin-1 gene knockout mice were successfully identified; reproduction of different mating methods The results basically accorded with Mendelian genetic law, and female and male caveolin-1-/- homozygous mice had certain reproductive ability, and the appearance characteristics of the three different genotype mice had no obvious difference.

　　Conclusion: The boiling lysis method to extract genomic DNA and PCR method can quickly and reliably identify the genotype of caveolin-1 gene knockout mice; the breeding method combining caveolin-1 heterozygous mice and homozygous mutual crossing may be short-term A better way to get enough homozygous offspring and homologous wild offspring.