Najjar S M, Lewis R E
Department of Pharmacology and Therapeutics, Medical College of Ohio, 3035 Arlington Avenue, Toledo, OH 43614, USA.
Gene. 1999 Apr 1;230(1):41-5. doi: 10.1016/s0378-1119(99)00052-9.
We have optimized a liposome-based transfection method that mediated highly efficient stable expression of foreign genes in hepatocytes. Moreover, we have observed that the metallothionein 1 promoter in the bovine papilloma virus-based expression vector drove the highest expression of foreign genes in hepatocytes as compared with the cytomegalovirus and the human polypeptide chain elongation factor 1alpha (EF-1alpha) promoters in the pcDNA 3-based expression vector. The cytomegalovirus promoter failed to yield significant expression in these cells. Furthermore, expression of foreign genes persisted up to at least 15 passages when expression was under the control of either the EF-1alpha or the metallothionein 1 promoter. Thus, these two promoters led to comparable stability of foreign genes in hepatocytes, with the metallothionein 1 promoter yielding a higher level of expression of foreign genes in these cells.
我们优化了一种基于脂质体的转染方法，该方法可介导外源基因在肝细胞中高效稳定表达。此外，我们观察到，与基于pcDNA 3的表达载体中的巨细胞病毒启动子和人多肽链延伸因子1α（EF-1α）启动子相比，基于牛乳头瘤病毒的表达载体中的金属硫蛋白1启动子在肝细胞中驱动外源基因的表达水平最高。巨细胞病毒启动子在这些细胞中未能产生显著表达。此外，当外源基因的表达受EF-1α或金属硫蛋白1启动子控制时，其表达至少持续15代。因此，这两个启动子在肝细胞中导致外源基因具有相当的稳定性，其中金属硫蛋白1启动子使这些细胞中外源基因的表达水平更高。
