表达小鼠Notch1胞内段基因慢病毒载体的构建及鉴定  

Construction and verification of lentivirus vector expressing mouse notch intracellular domain

作　　者：唐泉[1] 杨冰[1] 周慧[2] 杨福军[1] 段蓉[3] 樊体强[1] 韩英[1] 张晓东[1] 孙元明[1] 

机构地区：[1]中国医学科学院北京协和医学院放射医学研究所,天津分子核医学重点实验室,天津300192 [2]天津医科大学药学院,300070 [3]天津医科大学总医院药剂科,300052

出　　处：《国际生物医学工程杂志》2012年第3期133-136,I0002,共5页International Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目（30970867）;中国医学科学院放射医学研究所基金项目（SF1105,ST1223）

摘　　要：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.Objective Notch signaling is highly conservative in evolution and plays an important role in cell＇s proliferation and differentiation. Construction of lentiviral 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.

关 键 词：慢病毒 Notch胞内段 NOTCH1 

分 类 号：Q[生物学]