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目的:获得表达小鼠ROG(repressor of GATA 3)基因的重组腺相关病毒载体,制备表达ROG的重组腺相关病毒。方法:设计、合成引物,通过RT PCR法从CTLL 2细胞总RNA中扩增ROG基因,测序确认后,连接于pAAV MCS载体,双酶切、PCR鉴定重组病毒载体;制备高滴度重组腺相关病毒。结果:成功扩增了小鼠ROG基因,构建可表达ROG基因的重组腺相关病毒载体,并获得表达ROG的重组腺相关病毒。结论:本实验获得的小鼠ROG重组腺相关病毒,将为支气管哮喘的基因治疗研究奠定基础。
OBJECTIVE: To obtain a recombinant adeno-associated virus vector expressing mouse ROG (repressor of GATA 3) gene and prepare a recombinant adeno-associated virus expressing ROG. Methods: The ROG gene was amplified from total RNA of CTLL-2 cells by RT-PCR and sequenced. The ROG gene was ligated to pAAV MCS vector and double-digested. The recombinant adenovirus was identified by PCR. High-titer recombinant adeno-associated virus. Results: The mouse ROG gene was successfully amplified, a recombinant adeno-associated virus vector expressing ROG gene was constructed, and a recombinant adeno-associated virus expressing ROG was obtained. Conclusion: The mouse ROG recombinant adeno-associated virus obtained in this study will lay the foundation for the gene therapy of bronchial asthma.