动物造模,基因敲除 z-bio 2022-03-16 700

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

　　Methods: The imported caveolin-1 knockout mice were raised in SPF laboratory, and the genomic DNA of rat tail tissue was extracted by boiling lysis method, and the genotype was detected by PCR reaction according to the primer sequences provided by the Jackson Laboratory in the United States. , using caveolin-1+/- heterozygote cross, heterozygous and caveolin-1-/- homozygous cross (orthogonal and reverse cross), homozygous cross cross 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 knockout mice were successfully identified. The results basically conformed to the Mendelian inheritance law, and the female and male caveolin-1-/- homozygous mice had certain reproductive ability, and there was no significant difference in the appearance characteristics of the three different genotypes.

　　CONCLUSION: Genomic DNA extracted by boiling lysis method and PCR method can quickly and reliably identify the genotype of caveolin-1 knockout mice; the breeding method combining caveolin-1 heterozygous mouse cross and homozygous cross may be a short-term method. A better way to obtain sufficient numbers of homozygous mice and homologous wild mice.