摘要
指出了Kcnk2在多种疾病的发生机制中有着重要的地位,为了研究Kcnk2在相关疾病中的作用,利用所属CRISPR/Cas9系统的pL-CRISPR.EFS.GFP质粒,使用CRISPR设计工具设计敲除靶点,通过质粒酶切、引物退火、连接重组质粒,成功构建了Kcnk2的敲除载体,并通过DNA测序确定了插入序列。
Kcnk2 has an important role in a variety of diseases.In order to study the role Kcnk2 plays in those diseases, the CRISPR/Cas9 plasmid pL-CRISPR.EFS.GFP was used to construct the Kcnk2-knockout vector. The gRNA targeting sequence was designed by using CRISPR Design tools. By enzyme digestion, oligo annealing, and ligation to recombinant plasmid, Kcnk2 knockout vector was successfully constructed and confirmed by DNA sequencing.
作者
艾慧婷
刘欣煜
雷维琼
陈恒玲
Ai Huiting;Liu Xinyu;Lei Weiqiong;Chen Hengling(Biomedical Engineering College of South-Central University for Nationalities, Hubei Key Laboratoryof Medical Information Analysis and Tumor Diagnosis & Treatment, Wuhan, Hubei, 430074, China;Wuhan No.4 High School, Wuhan, Hubei, 430034, China)
出处
《绿色科技》
2019年第8期237-239,246,共4页
Journal of Green Science and Technology
