CAJ | 학술논문

目的 Notch信号通路在进化中高度保守,在细胞的生长和分化方面起重要作用.构建小鼠Notch胞内段(NICD)慢病毒载体,为Notch信号通路的研究奠定了基础.方法从C57BL/6J小鼠脊髓组织提取总RNA,逆转录合成cDNA,用PCR方法获取NICD的基因序列,构建慢病毒表达NICD载体,感染HEK293T细胞,实时定量PCR和Western blot分析NICD基因在靶细胞的表达.结果NICD基因在靶细胞的表达水平升高.结论成功构建表达小鼠NICD基因的慢病毒载体,为今后相关实验研究奠定了基础.
목적 Notch신호통로재진화중고도보수,재세포적생장화분화방면기중요작용.구건소서Notch포내단(NICD)만병독재체,위Notch신호통로적연구전정료기출.방법 종C57BL/6J소서척수조직제취총RNA,역전록합성cDNA,용PCR방법획취NICD적기인서렬,구건만병독표체NICD재체,감염HEK293T세포,실시정량PCR화Western blot분석NICD기인재파세포적표체.결과NICD기인재파세포적표체수평승고.결론 성공구건표체소서NICD기인적만병독재체,위금후상관실험연구전정료기출.
Objective Notch signaling is highly conservative in evolution and plays an important role in cell's proliferation and differentiation.Construction of tentiviral vector containing Notch intracellular domain (NICD) would lay the foundation for the study of Notch signaling.Methods Total RNA was extracted from the myeloid tissue of C57BL/6 mice.cDNA was composed via reverse transcription.NICD sequence was obtained by PCR and recombined into lentiviral vector.Lentiviral vector with NICD was infected into target HEK293T cell.Real-time PCR and Western blot were used to examine NICD expression in HEK293T cell.Results NICD expression increased in HEK293T cells.Conclusion The successful construction of lentiviral vector involving mice NICD expression provides the foundation for the future study.