FEEDING, BREEDING, AND IDENTIFYING UCH-L1 GENE KNOCKOUT MICE
Objective To obtain UCH-L1 gene knockout homozygous mice by feeding,breeding,and identifiying ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) knockout mice,so as to lay the foundation for further study of the biological function of UCH-L1.Methods Seven UCH-L1 gene knockout mice purchased from the United States were fed for one week to adapt new circumstance,then one female heterozygous mouse and one male heterozygous mice were put into a cage for mating and breeding.The tissue of tail end (0.5 cm in length) of the offspring was clipped to extract the genomic DNA,and PCR method was used for identification of the genotype.The offspring of wild-type(UCH-L1+/+),and UCH-L1 gene knockout homozygous(UCH-L1-/-) mice were used for subsequent biological function study;the heterozygous (UCH-L1 +/-) mice for breeding.The phenotypic changes of UCH-L1 gene knockout homozygous mice were observed,and the UCH-L1 gene knockout effect was verified by Western blotting.Results Mice purchased from abroad was successfully reproduced.A total of 370 filial generation mice were reproduced;and genotypic identification revealed that the percentage of UCH-L1+/+ mice was 34.1％,of UCH-L1+/-mice 50.5％,and of UCH-L1 / mice 15.4％.UCH-L1 gene knockout homozygous mice were obtained successfully.The methods for feeding,breeding,and identification of UCH-L1 gene knockout mice were established.Conclusion The usage of male and female UCH-L1 heterozygous mice for mating,not only can get UCH-L1 gene knockout homozygous mice,but also can conduce to the long-term feeding and breeding.
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NSTL
万方数据
维普
