CAJ | 학술논문

目的探讨GJB2基因条件敲除（conditional connexin 26 knock out ，cCx26KO）小鼠的饲养、繁殖及基因型鉴定方法，为进一步研究GJB2基因突变导致非综合征型聋的机制奠定基础。方法将引进的两对转基因小鼠Cx26loxp/loxp和Pax2-Cre/＋进行交配饲养与繁殖，选取子一代雌性Cx26loxp/-＿Pax2-Cre/＋小鼠与雄性小鼠Cx26loxp/loxp合笼交配，即获得cCx26KO小鼠。提取鼠尾组织基因组DNA ，PCR方法鉴定动物基因型。采用c-ABR检测成年cCx26KO小鼠和野生型小鼠的听力，进一步验证PCR方法的正确性。结果 cCx26KO小鼠繁殖成功后，采用 PCR 方法鉴定分析，成功获得 Cx26loxp/loxp＿Pax2Cre/＋、Cx26loxp/-＿Pax2- Cre/＋、Cx26loxp/loxp、Cx26loxp/-4种基因型小鼠，其繁殖结果符合孟德尔遗传定律。与野生型小鼠相比，cCx26KO小鼠c-ABR反应阈显著升高，约达95 dB SPL。结论 PCR方法可准确鉴定子鼠的基因型，雌性 Cx26loxp/-＿Pax2-Cre/＋小鼠与雄性Cx26loxp/loxp小鼠交配是获得cCx26KO实验小鼠的有效途径。
목적탐토GJB2기인조건고제（conditional connexin 26 knock out ，cCx26KO）소서적사양、번식급기인형감정방법，위진일보연구GJB2기인돌변도치비종합정형롱적궤제전정기출。방법장인진적량대전기인소서Cx26loxp/loxp화Pax2-Cre/＋진행교배사양여번식，선취자일대자성Cx26loxp/-＿Pax2-Cre/＋소서여웅성소서Cx26loxp/loxp합롱교배，즉획득cCx26KO소서。제취서미조직기인조DNA ，PCR방법감정동물기인형。채용c-ABR검측성년cCx26KO소서화야생형소서적은력，진일보험증PCR방법적정학성。결과 cCx26KO소서번식성공후，채용 PCR 방법감정분석，성공획득 Cx26loxp/loxp＿Pax2Cre/＋、Cx26loxp/-＿Pax2- Cre/＋、Cx26loxp/loxp、Cx26loxp/-4충기인형소서，기번식결과부합맹덕이유전정률。여야생형소서상비，cCx26KO소서c-ABR반응역현저승고，약체95 dB SPL。결론 PCR방법가준학감정자서적기인형，자성 Cx26loxp/-＿Pax2-Cre/＋소서여웅성Cx26loxp/loxp소서교배시획득cCx26KO실험소서적유효도경。
Objective To explore the methods of breeding ,reproduction and genotype of GJB2 knock -out (cCx26KO) mice and further study the critical role of GJB2 mutation in the onset of nonsyndromic hearing loss (NSHL) .Methods Two pairs of transgenic mice (Cx26loxp/loxp and Pax2 -Cre/+ ) were inbreeded to produce Cx26loxp/-_Pax2-Cre/+ ones ,female of which were used to mate with the male Cx26loxp/loxp ones to finally get the Cx26loxp/loxp_Pax2 -Cre/+ mice(cCx26KO) .The genotype was done by PCR and Agarose gel electro-phoresis using genome DNA extracted from the mice tails .The c-ABR was used to detect the hearing ability of the cCx26KO mice .Results Both breeding and reproduction of cCx26KO mice were successful .It was fruitful to obtain four genotype mice(Cx26loxp/loxp_ Pax2-Cre / + ,Cx26loxp / -_Pax2-Cre / + ,Cx26loxp/loxp ,Cx26loxp /-) by the breeding Cx26loxp / -_Pax2Cre / + and Cx26loxp/loxp mice .The results of breeding were met with the Mendel's law .The c-ABR revealed elevated response threshold around 95 dB SPL in cCx26KO mouse compared to the wild type ones ,which further validated the accuracy of the PCR method .Conclusion The PCR method is cor-rectly identified sub pups genotype and the female Cx26loxp/-_Pax2-Cre/+ mice mating with the male Cx26loxp/loxp ones is an effective way to obtain the cCx 26KO mice .