PD-1人源化小鼠构建繁殖与基因型鉴定

Breeding and Genotype Identification of PD-1 Humanized Mice

目的探讨PD-1人源化小鼠的繁育与基因型鉴定,为进一步研究相关小分子抑制剂的体内药效评价提供动物模型。方法通过构建PD-1人源化小鼠,获得F1代鼠4只,雌雄交配进行培育繁殖。对每窝仔鼠的数量,存活率等进行记录观察,随后对仔鼠剪尾提取基因组DNA,用PCR法扩增目的基因,然后进行核酸电泳,鉴定基因型。选用F2代纯合型与纯合型雌雄交配,野生型与野生型雌雄交配。结果鉴定的F2代小鼠有野生型、纯合型和杂合型3种基因型。纯合型与纯合型交配获得F3代仔鼠基因型全部为纯合型。野生型与野生型交配获得F3代仔鼠基因型全部为野生型。结论成功筛选出PD-1人源化小鼠纯合子基因型,并有效地进行扩群保种,为今后应用该小鼠模型提供实验保障。

Objective To explore the breeding and genotype identification of PD-1 humanized mice,and provide an animal model for further research on the efficacy evaluation of small molecule inhibitors in vivo.Method Through the construction of PD-1 humanized mice,four F1 generation mice were obtained,which were bred by mating male and female.The number and survival rate of each litter of mice were recorded and observed,and then genomic DNA was extracted from the offspring by tail clipping,the target gene fragment was amplified by PCR,and then the genotype was identified by nucleic acid electrophoresis.The F2 generation homozygous type and homozygous type were used for male and female mating,and wild type and wild type were used for male and female mating.Result The F2 generation mice were identified as wild type,homozygous type and heterozygous type.The genotype of F3 offspring obtained from homozygous and homozygous mating were all homozygous.The genotype of F3 offspring obtained from wild type and wild type mating were all wild type.Conclusion The homozygous genotype of PD-1 humanized mouse was successfully screened,and the population expansion and seed conservation were effectively carried out,providing experimental guarantee for the future application of this mouse model.