免疫蛋白酶体亚基低分子重量蛋白2基因敲除大鼠的繁殖与基因型鉴定  

Breeding and identifying for immunoproteasome subunit low molecular weight protein 2 gene knockout rats

作　　者：陈兴泳[1,2] 毛艳光 姚楠楠 汪银洲 郭越婷[3] 刘迎春 CHEN Xing-yong;MAO Yan-guang;YAO Nan-nan;WANG Yin-zhou;GUO Yue-ting;LIU Ying-chun(Department of Neurology,Fujian Provincial Hospital,Fujian Medical University Provincial Clinical Medical College,Fuzhou 350001,Fujian Province,China;Fujian Provincial Key Laboratory of Critical Care Medicine,Fuzhou 350001,Fujian Province,China;Key Laboratory of Stem Cell Engineering and Regenerative Medicine,School of Basic Medical Sciences,Fujian Medical University,Fuzhou 350122,Fujian Province,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院神经内科,福建福州350001 [2]福建省疑难重症研究重点实验室,福建福州350001 [3]福建医科大学基础医学院干细胞工程与再生医学重点实验室,福建福州350122

出　　处：《中国临床药理学杂志》2022年第21期2610-2614,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81771250);福建省医学创新课题基金资助项目(2021CXB002);福建省自然科学基金资助项目(2020J011059,2020R1011004);福建省立医院“创双高”火石基金资助项目(2020HSJJ07)。

摘　　要：目的进行完全性敲除免疫蛋白酶体亚基低分子重量蛋白2(LMP2)基因(LMP2^(-/-))Sprague-Dawley(SD)大鼠繁殖及鉴定。方法LMP2基因杂合子(LMP2^(+/-))SD大鼠饲养及配种繁殖,提取子代大鼠鼠尾基因组DNA,采用聚合酶链反应(PCR)方法进行基因型鉴定,将获得的纯合子大鼠与异性杂合子交配,以获得更多的基因敲除纯合子大鼠。结果LMP2杂合子SD大鼠繁殖生成子代有37只,日常饮食、活动和生长发育在各动物间未见无明显差异。37只子代大鼠基因型分别为野生型(WT)(LMP2^(+/+))9只、杂合子型(LMP2^(+/-))17只和纯合子型(LMP2^(-/-))11只。各基因型的PCR产物经琼脂糖凝胶电泳条带情况分别为野生型未出现条带,纯合子大鼠仅出现500bp条带,杂合子大鼠出现500 bp、738 bp两个条带。结论LMP2基因敲除大鼠成功繁殖,为后续实验奠定了基础。Objective To breed,reproduce and identify for immunoproteasome subunit low molecular weight protein 2(LMP2)gene completed knockout(LMP2^(-/-))Sprague-Dawley(SD)rats.Methods LMP2 heterozygous(LMP2^(+/-))rats were bred and mated.The tail genomic DNA of offspring rats were extracted and identified by polymerase chain reaction(PCR).The obtained homozygous rats were mated with the opposite sex heterozygous rats to obtain more homozygous rats with knockout LMP2 gene.Results There were 37 progenies generated from LMP2^(+/-)SD rats,and there were no significant differences in diet,activity,growth and development among the animals.The genotypes of 37 progenies were wild type(WT)(LMP2^(+/+))9,heterozygous(LMP2^(+/-))17 and homozygous(LMP2^(-/-))11,respectively.The products by PCR amplification and agarose gel electrophoresis indicated that the wild type had no bands,only 500 bp bands were found in homozygous rats,and 500 bp and 738 bp bands were found in heterozygous rats.Conclusion LMP2 gene knockout rats successfully propagated,which laid a foundation for the subsequent experiments.

关 键 词：基因敲除大鼠 免疫蛋白酶体 纯合子 杂合子 

分 类 号：R97[医药卫生—药品]