白细胞介素1受体1和白细胞介素6双基因敲除小鼠的构建及基因型鉴定  

Construction and genotype identification of IL-1R1-/-IL-6-/-gene mice

作　　者：范海涛 李恒 郭磊[1] 王晶晶[1] 储曼曼 黄星 郑惠文[1] 杨泽宁[1] 李洪哲[1] 刘龙丁[1] Fan Haitao;Li Heng;Guo Lei;Wang Jinxing;Chu Manman;Huang Xing;Zheng Huiwen;Yang Zening;Li hongzhe;Liu Longding(Institue of Medical Biology,Chinese Academy of Medical Sciences ＆ Peking Union Medical College,Kunming 650031,China;Kunming Medical University,Kunning 650031,Chin)

机构地区：[1]中国医学科学院,北京协和医学院医学生物学研究所,昆明650031 [2]昆明医科大学,650031

出　　处：《国际免疫学杂志》2018年第3期251-255,共5页International Journal of Immunology

基　　金：国家自然科学基金（81373142）;云南省科技创新人才计划（2016HC009）

摘　　要：目的通过白细胞介素1受体1 （interleukin-1 receptor1, IL-1R1）和白细胞介素6 （interleukin-6，IL-6）单基因敲除小鼠杂交繁育以获得IL-1R1-/-IL-6-/-纯合子小鼠。方法将IL-1R1＋/＋IL-6-/-和IL-1R1-/-IL-6＋/＋小鼠杂交，并进行子代自交繁殖，提取基因组DNA，PCR鉴定其自交子代基因型。并用免疫印迹法（Western blot）、酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）确定自交小鼠后代基因型。 结果PCR鉴定成功获得基因型为IL-1R1-/-IL-6-/-的双基因敲除小鼠。Western blot结果显示，双基因敲除小鼠未见IL-1R1的蛋白条带；ELISA结果提示，双基因敲除小鼠的IL-6细胞因子为阴性。结论成功获得IL-1R1-/-IL-6-/-双基因敲除纯合小鼠，为深入研究IL-1R1、IL-6基因相关的炎症疾病动物模型奠定基础。ObjectiveTo construct mice with the knockout of both interleukin 1 receptor 1 （IL-1R1） and interleukin 6 （IL-6） genes.MethodsWe generated the IL-1R1＋ /-IL-6＋ /-offspring by crossing the IL-1R1＋ /＋ mice with IL-6＋ /＋ mice.DNA was extracted from the ear tissue of mouse offspring fand and subjected to PCR for genotype identification.Mice with IL-1R1-/-IL-6-/-were identified.Mice with the genotype of IL-1R1-/-IL-6-/-were further confirmed by the analysis of Western blotting and enzyme-linked immunosorbent assay（ELISA）. ResultsTwo mice with genotype of IL-1R1-/-IL-6-/-were obtained. Western blot and ELISA analysis showed IL-1R1 and IL-6 were not expressed in IL-1R1-/-IL-6-/-mice.ConclusionWe have successfully established IL-1R1-/-IL-6-/-gene-knockout mice.This work would provide foundation for the further study of the inflammatory diseases related to IL-1R1 and IL-6.

关 键 词：白细胞介素-1受体1 白细胞介素-6 基因敲除小鼠 基因型鉴定 

分 类 号：R392[医药卫生—免疫学] R-332[医药卫生—基础医学]