花生FAD2基因CRISPR/Cas9多靶点敲除载体构建  

Construction of CRISPR/Cas9 Multi-Target Knockout Vector of FAD2 Gene in Peanut

作　　者：李柯 赵昆昆 宁龙龙 马倩 李忠峰 马兴立[1] 张幸果[1] 殷冬梅[1] Li Ke;Zhao Kunkun;Ning Longlong;Ma Qian;Li Zhongfeng;Ma Xingli;Zhang Xingguo;Yin Dongmei(College of Agronomy,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学农学院

出　　处：《山东农业科学》2019年第9期56-62,67,共8页Shandong Agricultural Sciences

基　　金：国家自然科学基金项目(U1704232);河南省产业技术体系项目(S2012-05-G03)

摘　　要：ω-6脂肪酸脱氢酶控制花生中油酸向亚油酸的转化,该酶由AhFAD2基因编码。为了研究AhFAD2基因的功能,本研究根据前期已经克隆得到的AhFAD2基因序列,设计了gRNA前导序列,并构建了CRISPR/Cas9基因编辑载体,通过双酶切和Sanger测序确定载体构建成功。AhFAD2基因CRISPR/Cas9载体的构建为接下来的遗传转化及研究该基因的功能奠定了基础。The omega-6 fatty acid dehydrogenase controls the conversion of oleic acid into linoleic acid in peanut, which is encoded by the AhFAD2 gene. In order to research the function of AhFAD2 gene, the gRNA leader sequence was designed based on the sequence of AhFAD2 gene cloned in previous stage, and the CRISPR/Cas9 gene editing vector was constructed in this study. The vector was successfully constructed after determined by double enzyme digestion and Sanger sequencing. The construction of AhFAD2 gene CRISPR/Cas9 vector laid foundations for the subsequent genetic transformation and function study of the gene.

关 键 词：花生 FAD2 CRISPR/Cas9 基因敲除 

分 类 号：S565.2[农业科学—作物学] Q782[生物学—分子生物学]