CAJ | 학술논문

目的：核干细胞因子( nucleostemin,NS)是维持干细胞和癌细胞增殖所必需的蛋白质,可能成为肿瘤基因治疗的潜在靶点。本文旨在构建靶向NS干扰载体pGenesil-1-NS,为后续实验奠定基础。方法基于已发布的NS mRNA序列(NM_206825),选取5′-AAGCCTA GGAAAGACCCAGG-3′(397-416)作为候选靶序列,按shRNA载体设计原则,设计合成两条互补DNA链,退火后插入载体,并以酶切和测序鉴定重组转化子。结果经酶切及测序鉴定证实合成序列正确插入载体。结论成功构建NS特异性干扰载体pGenesil-1-NS,可用于NS在肿瘤中的功能研究。
목적：핵간세포인자( nucleostemin,NS)시유지간세포화암세포증식소필수적단백질,가능성위종류기인치료적잠재파점。본문지재구건파향NS간우재체pGenesil-1-NS,위후속실험전정기출。방법기우이발포적NS mRNA서렬(NM_206825),선취5′-AAGCCTA GGAAAGACCCAGG-3′(397-416)작위후선파서렬,안shRNA재체설계원칙,설계합성량조호보DNA련,퇴화후삽입재체,병이매절화측서감정중조전화자。결과경매절급측서감정증실합성서렬정학삽입재체。결론성공구건NS특이성간우재체pGenesil-1-NS,가용우NS재종류중적공능연구。
Objective Nucleostemin,a protein to maintain the proliferations of stem cell and cancer cell, may become a potential target for tumor gene therapy. This study aimed to construct the NS specific interference vector pGenesil-1-NS,and to lay a foundation for further study. Method Based on the published NS-mRNA sequence (NM_206825),5′-AAGCCTA GGAAAGACCCAGG-3′(397-416) was selected as candidate target sequence. According to the shRNA carrier design principle,shDNA that could be transcripted to shRNA was designed and synthesized,then inserted into pGenesil-1 vector. The recombinant was identified by restriction enzyme digestion analysis followed by sequence analysis. Results Confirmed by restriction analysis and sequencing,shDNA was correctly inserted into the vector. Conclusion NS-target interference vector pGenesil-1-NS was successfully constructed,which would be applied in the study of NS function in tumor generation and progression.