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 UCHL1 gene knockout homozygous mice,but also can conduce to the long-term feeding and breeding.