摘要
目的检测真核表达质粒载体pIRES2-EGFP-NT3、pIRES2-DsRed2-Hath1在阳离子脂质体介导下对HEK293T细胞的共转染情况。方法分别提取含有目的基因NT3的真核表达质粒载体pIRES2-EGFP-NT3和含有目的基因Hath1的真核表达质粒载体pIRES2-DsRed2-Hath1,在阳离子脂质体LipofectamineTM2000介导下将其共转染HEK293T细胞,转染后24h,通过Confocal显微镜观察HEK293T细胞的转染情况和转染效率。结果 pIRES2-EGFP-NT3和pIRES2-DsRed2-Hath1能有效转染并共转染HEK293T细胞,其转染率pIRES2-EGFP-NT3为23.79%(157/660);pIRES2-DsRed2-Hath1为8.03%(53/660);共转染率为3.64%(24/660)。结论 pIRES2-EGFP-NT3和pIRES2-DsRed2-Hath1在阳离子脂质体介导下对HEK293T细胞的共转染成功为NT3和Hath1基因共转染耳蜗的实验奠定基础。
Objective To detect the co-transfected capability of pIRES2-EGFP-NT3 and pIRES2- DsRed2-Hathl in HEK293T by cationic hposome. Methods After pIRES2-EGFP-NT3 and pIRES2- DsRed 2 - Hath 1 had been abstracted successfully, HEK 293 T was co - transfected with pIRES 2 - EGFP - NT 3 and pIRES2- DsRed2- Hath 1 by Lipofectamine^TM 2000. Twenty four hours later, the efficiency of the transfection was analyzed with Confoeal microscope. Results The pIRES 2 - DsRed 2 - Hath 1 and pIRES 2 - EGFP - NT 3 could be effectively co-transfected into HEK293 T by cationic liposome： 157 out of 660 cells were transfected by pIRES2-EGFP-NT3 , 53 out of 660 cells by pIRES2-DsRed2-Hathl , 24 out of 660 cells by pIRES2- EGFP-NT3 and pIRES2-DsRed2-Hathl. Conclusion The effective co-transfection of pIRES2-EGFP-NT3 and pIRES 2 - DsRed 2 - Hath 1 into HEK 293 T by Lipofectamine^TM 2000 lays the basis for further study, such as co- transfection of NT3 and Hath 1 gene in deaf cochlea, and so on.
出处
《中国耳鼻咽喉颅底外科杂志》
CAS
2010年第5期335-340,共6页
Chinese Journal of Otorhinolaryngology-skull Base Surgery
基金
江苏省医学重点人才项目(RC2007010)
南京市医学重点发展项目(ZKX06019)
关键词
神经营养因子3
质粒
脂质体
实验研究
Neurotrophin 3
Plasmid
Cationic liposome
Empirical study
