| 利用CRISPR/Cas9系统构建Prdx6基因敲除的RAW264.7细胞系 |
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| 引用本文： | 翟菲菲,全芷仪,王路鹿,张士军,鞠丹迪,水一鸣,常江,常恒祯,胡盼,李岩松,卢士英,周玉,柳增善,李兆辉,任洪林.利用CRISPR/Cas9系统构建Prdx6基因敲除的RAW264.7细胞系[J].动物医学进展,2019,40(4):36-40. |
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| 作者姓名： | 翟菲菲 全芷仪 王路鹿 张士军 鞠丹迪 水一鸣 常江 常恒祯 胡盼 李岩松 卢士英 周玉 柳增善 李兆辉 任洪林 |
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| 作者单位： | 吉林大学动物医学学院人兽共患病研究所/人兽共患病研究教育部重点实验室 ,吉林长春,130062;长春市实验中学 ,吉林长春,130000 |
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| 基金项目： | 国家重点研发计划;吉林省科技发展计划 |
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| 摘 要： | 利用CRISPR/Cas9系统构建敲除Prdx6基因的RAW264.7小鼠巨噬细胞系,为探讨Prdx6与布鲁菌感染及胞内寄生的关系构建细胞模型。针对Prdx6基因的第1外显子设计sgRNA,构建重组表达载体Prdx6-pX459-sgRNA,将重组质粒转染RAW264.7细胞,嘌呤霉素初步筛选。单细胞接种并扩繁,提取基因组DNA,PCR及测序鉴定。结果表明,在敲除细胞株基因组中存在碱基缺失,导致移码突变。说明成功获得敲除Prdx6基因的RAW264.7细胞系。
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| 关 键 词： | CRISPR/Cas9 RAW264.7 Prdx6 移码突变 布鲁菌 |
Construction of Prdx6-Knockout Raw264.7 Cell Line by CRISPR/Cas9 System |
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| ZHAI Fei-fei,QUAN Zhi-yi,WANG Lu-lu,ZHANG Shi-jun,JU Dan-di,SHUI Yi-ming,CHANG Jiang,CHANG Heng-zhen,HU Pan,LI Yan-song,LU Shi-ying,ZHOU Yu,LIU Zeng-shan,LI Zhao-hui,REN Hong-lin.Construction of Prdx6-Knockout Raw264.7 Cell Line by CRISPR/Cas9 System[J].Progress In Veterinary Medicine,2019,40(4):36-40. |
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| Authors: | ZHAI Fei-fei QUAN Zhi-yi WANG Lu-lu ZHANG Shi-jun JU Dan-di SHUI Yi-ming CHANG Jiang CHANG Heng-zhen HU Pan LI Yan-song LU Shi-ying ZHOU Yu LIU Zeng-shan LI Zhao-hui REN Hong-lin |
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| Institution: | (College of Veterinary Medicine,Institute of Zoonosis,Key Laboratory of Zoonosis Research Ministry of Education,Jilin University,Changchun Jilin,130062,China;Changchun City Experimental High School,Changchun,Jilin,130000,China) |
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| Abstract: | ZHAI Fei-fei;QUAN Zhi-yi;WANG Lu-lu;ZHANG Shi-jun;JU Dan-di;SHUI Yi-ming;CHANG Jiang;CHANG Heng-zhen;HU Pan;LI Yan-song;LU Shi-ying;ZHOU Yu;LIU Zeng-shan;LI Zhao-hui;REN Hong-lin(College of Veterinary Medicine,Institute of Zoonosis,Key Laboratory of Zoonosis Research Ministry of Education,Jilin University,Changchun Jilin,130062,China;Changchun City Experimental High School,Changchun,Jilin,130000,China) |
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| Keywords: | CRISPR/Cas9 RAW264 7 peroxiredoxin 6 frameshift mutation Brucella |
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