Fam172a基因敲除小鼠的制备与基因型鉴定  

Preparation and genotype identification of Fam172a gene knockout mice

作　　者：张一帆[1] 王建文[2] 杨琪[1] 郝晓花[3] 黄玉波[3] 魏红山[1,2] 

机构地区：[1]北京大学地坛医院教学医院消化科,北京100015 [2]首都医科大学附属北京地坛医院消化科,北京100015 [3]首都医科大学附属北京地坛医院肝病中心,北京100015

出　　处：《中国肝脏病杂志（电子版）》2017年第2期32-35,共4页Chinese Journal of Liver Diseases：Electronic Version

基　　金：国家自然科学基金项目(81271901);北京市自然科学基金项目(7152073)

摘　　要：目的建立稳定的Fam172a基因敲除C57/B6J小鼠模型,观察基因敲除对小鼠生长发育的影响以及该基因在小鼠体内的生物学的作用。方法采用基因打靶敲除技术,获得杂合子(Fam172a^(+/-))小鼠。将所得Fam172a^(+/-)小鼠饲养于屏障环境中杂交获得纯合子(Fam172a^(-/-))小鼠。利用聚合酶链式反应(polymerase chain reaction,PCR)技术对小鼠基因型进行鉴定,称量出生后20天、40天以及60天野生型(wild type,WT)小鼠及Fam172a^(-/-)小、鼠的体重。随机处死8周龄WT小鼠和Fam172a^(-/-)小鼠,取心、肝、脾、肺及肾组织,采用Western blot险测Fam172a基因的组织表达特异性。结果 PCR及Western blot结果均证明Fam172a基因在Fam172a^(-/-)小鼠体内完全删除。Farm172a^(-/-)小鼠与WT小鼠20天、40天以及60天体重的差异均有统计学意义(P均<0.05)。FAM172A蛋白在WT小鼠中的表达有组织特异性。Fam172a^(-/-)小鼠每胎产仔数略低于WT小鼠,差异无统计学意义(P>0.05)。各小鼠外观、行为和繁殖力无显著差异。结论 Fam172a基因敲除C57/B6J小鼠模型已成功构建,并繁育出足够数量的Fam172a^(-/-)小鼠进行下一步实验。Fam172a基因对小鼠体重和生长发育具有重要作用。Objective A Fam172 a gene knockout C57/B6 J mice model was generated to explore the functions of Fam172 a gene and examine mechanisms of its growth and development in vivo.Methods Total of six Fam172a＋/- mouse were got by gene targeting.The Fam172a-/- mouse were successfully produced by hybridization of Fam172a＋/- mice and bred in a barrier environment.The mouse genotypes were identified by PCR.The body weights of WT and Fam172a-/- mouse were measured at 20,40 and 60 days after birth.Eightweek-old male WT and Fam172 a gene knockout mouse were randomly selected and sacrificed,and their heart,liver,spleen,lung and renal were removed and used for Western blot to analyze the expression of Fam172 a gene at indicated tissues.Results The results of PCR and Western blot showed that Fam172 a gene was totally knocked out in Fam172a-/- mice.The body weights of WT and Fam172a-/- mouse were significantly different at 20,40 and 60 days after birth（P〈0.05）.The expression of FAM172 A protein in WT mice had tissue specificity.Litter sizes of Fam172a-/- mouse were slightly lower than those of WT mouse,and had no statistical difference（P〉0.05）.There was no significant difference on the appearances,behaviors and reproductive performance.Conclusion Fam172a-/- mouse were prepared.This approach allows breeding of a sufficient number of Fam172 a gene knockout C57/B6 J mouse as the animal models for studying the role of Fam172 a gene.The data revealed that Fam172 a gene is critically required for body weight,growth and development of mouse.

关 键 词：Fam172a 基因敲除 小鼠 

分 类 号：Q78[生物学—分子生物学]