Liu Yu-kou, Chu Chih-chieh, Wu Tzong-yuan
Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan, Taiwan, China.
Acta Pharmacol Sin. 2006 Mar;27(3):321-7. doi: 10.1111/j.1745-7254.2006.00276.x.
To identify a shuttle promoter that can mediate gene expression in both insect cells and mammalian cells to facilitate the development of a baculovirus vector-based mammalian cell gene delivery vehicle.
Recombinant baculoviruses carrying the beta-galactosidase reporter gene under the control of an early to late (ETL) promoter of the Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) or a cytomegalovirus immediate early promoter (CMV promoter) were constructed. COS1, HeLa, CHO-K1, hFob1.19, and MCF-7 mammalian cells were tested for the expression of b-galactosidase.
ETL promoter activity was higher in bone-derived hFob1.19 than in COS1, HeLa, CHO-K1, or MCF-7 mammalian cells. The transient plasmid transfection assay indicated that ETL promoter activity in mammalian cells was dependent on baculovirus gene expression.
ETL promoter activity in mammalian cells is baculovirus gene expression-dependent, and the shuttle promoter will facilitate the application of baculovirus expression vectors in mammalian cell expression systems and for gene therapy.
鉴定一种能在昆虫细胞和哺乳动物细胞中均介导基因表达的穿梭启动子，以促进基于杆状病毒载体的哺乳动物细胞基因递送载体的开发。
构建了携带在苜蓿银纹夜蛾多核多角体病毒（AcMNPV）的早期至晚期（ETL）启动子或巨细胞病毒立即早期启动子（CMV启动子）控制下的β-半乳糖苷酶报告基因的重组杆状病毒。对COS1、HeLa、CHO-K1、hFob1.19和MCF-7哺乳动物细胞进行β-半乳糖苷酶表达检测。
ETL启动子活性在骨源hFob1.19细胞中高于COS1、HeLa、CHO-K1或MCF-7哺乳动物细胞。瞬时质粒转染试验表明，哺乳动物细胞中的ETL启动子活性依赖于杆状病毒基因表达。
哺乳动物细胞中的ETL启动子活性依赖于杆状病毒基因表达，该穿梭启动子将促进杆状病毒表达载体在哺乳动物细胞表达系统及基因治疗中的应用。
