Genotype identification and breeding method of caveolin-1 gene knockout mice
Received:December 11, 2015  
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KeyWord:Caveolin-1;Gene knockout;Mice;Identification;Breeding methods
周胜强 湖南中医药大学, 长沙
罗东 湖南中医药大学, 长沙
黄素芬 湖南中医药大学, 长沙
易健 湖南中医药大学第一附属医院, 长沙
刘柏炎 湖南中医药大学, 长沙
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      Objective To investigate the identification and optimal breeding method of caveolin-1 knockout mice, and provide an ideal animal model for further study of the role of caveolin-1 in cerebral ischemic injury and repair. Methods The introduced caveolin-1 gene knockout mice were reared in the SPF laboratory and genomic DNA was extracted from mouse tail tissue by the method of boiling lysis. According to the primer sequences provided by the Jackson Laboratory of America for polymerase chain reaction (PCR) to detect the genotypes, with the four different ways of mating:caveolin-1+/-heterozygote intercrossing, heterozygous and homozygous caveolin-1-/- hybrid (orthogonal and pay) as well as homozygous intercrossing. The pregnancy rate, shape characteristics of the filial generation mice and homozygous rate of the parental mice were observed. Results Agarose gel electrophoresis results indicated that the size of molecular weight of the PCR products was about 200 bp and 661 bp, which were consistent with the expected target gene fragment, and identified caveolin-1 gene knockout mice of different genotypes successfully. The results of different mating patterns are basically in agreement with Mendel rule, and the female and male aveolin-1-/- homozygous mice had a certain ability to reproduce, three different genotypes of mice had no significant differences between the shape features. Conclusions PCR can fast and reliably identify the genotypes of caveolin-1 knockout mice using genomic DNA through the method of boiling lysis. Combining the breeding methods of intercrossing of caveolin-1 heterozygous mice and intercrossing of caveolin-1 homozygous mice may be a good way to obtain enough homozygous mice and homologous wild type mice in a short period.
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