| 摘要: |
| 目的 繁殖及鉴定NLRP3基因敲除（NLRP3-/-）小鼠。方法 将引进的NLRP3基因敲除杂合子小鼠，单独进行饲养及配种繁殖，繁殖后其子代出现3种基因型：野生型、杂合子型及纯合子型，提取每只小鼠的基因组DNA，用PCR和T7E1酶切方法进行鉴定，将获得的纯合子小鼠与异性杂合子交配，以获得更多的基因敲除纯合子小鼠。结果 NLRP3杂合子小鼠的饲养及繁殖均获得成功，获得了NLRP3基因敲除纯合子和杂合子小鼠。结论 正确的饲养、繁殖及鉴定方法是从NLRP3基因敲除杂合子小鼠中获得纯合子小鼠的有效途径。 |
| 关键词: 基因敲除小鼠 NLRP3 炎症小体 纯合子 杂合子 |
| DOI：10.3724/SP.J.1008.2014.00555 |
| 投稿时间：2014-03-10修订日期：2014-04-13 |
| 基金项目:国家“重大新药创制”科技专项（2014ZX09J14106）；上海市公共卫生重点学科建设计划项目（12GWZX0501）；上海市自然科学基金（13ZR1449400）. |
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| Breeding and genotype identification of NLRP3 gene knock-out mice |
| FENG Shi-jie,ZONG Ying,ZHANG Zi-teng,DU Xiu-ming,YUAN Bo-jun,LU Guo-cai* |
(Department of Health Toxicology, College of Tropical Medicine and Public Health, Second Military Medical University, Shanghai 200433, China
*Corresponding authors.) |
| Abstract: |
| Objective To breed and identify NLRP3 gene knock-out mice. Methods The NLRP3 gene knock-out heterozygote mice were bred alone and copulated. The offsprings were to have three genotypes: wild genotype, heterozygote genotype and homozygote genotype. Genomic DNA was obtained from each pups and were subjected to PCR and T7 endonuclease 1 to identify the genotype. The homozygote mice were mated with the opposite sex heterozygote mice to obtain more homozygote pups. Results Breeding and reproducing were both successful, and we obtained heterozygote genotype and homozygote genotype mice with NLRP3 gene knock-out. Conclusion Correct methods of breeding, reproducing and identifying can effectively obtain NLRP3 gene knock-out mice from heterozygote mice. |
| Key words: knockout mice NLRP3 inflammasome mice homozygote heterozygote |
