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目的:构建小鼠Dnd1基因的RNAi慢病毒载体。方法:针对小鼠Dnd1基因RNAi设计有效靶序列,合成靶序列的Oligo DNA。退火形成双链DNA,与经Age I和EcoR I酶切后的pGCSIL-GFP载体连接产生短发卡RNA慢病毒载体,PCR筛选阳性克隆,测序鉴定。结果:PCR鉴定与DNA测序证实合成的含Dnd1sh RNA慢病毒载体寡核苷酸链插入正确。结论:成功构建小鼠Dnd1基因的RNAi慢病毒载体。
Objective: To construct RNAi lentiviral vector of mouse Dnd1 gene. Methods: Targeting the Dnd1 gene RNAi of mice to design effective target sequence and synthesizing target sequence Oligo DNA. Annealed to form double-stranded DNA, which was ligated with pGCSIL-GFP vector digested with Age I and EcoR I to generate short hairpin RNA lentivirus vector. Positive clones were screened by PCR and identified by sequencing. RESULTS: PCR and DNA sequencing confirmed the correct insertion of the synthetic DNA strand containing the Dnd1sh RNA lentivirus vector. Conclusion: RNAi lentiviral vector of mouse Dnd1 gene was successfully constructed.