牛白血病病毒(BLV)和巨细胞病毒启动子驱动的报告基因在BLV感染和未感染细胞中的表达比较

Jerome S Harms, Kurt A Eakle, Lillian S Kuo, Robert D Bremel, Gary A Splitter
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引用次数: 7

摘要

背景:病毒启动子被用于哺乳动物表达载体,因为它们通常在不同组织和物种的多种细胞中具有很强的活性。方法:通过与CMV启动子(CMVp)的直接比较,研究BLV LTR/启动子(BLVp)在哺乳动物表达载体中的应用。用BLVp或CMVp驱动的荧光素酶载体(包括D17、FLK、BL3.1和原代牛B细胞)稳定转导不同组织和物种的细胞系,利用荧光素酶测定启动子活性。通过添加BLV Tax表达载体和/或BLV感染以及曲古斯汀A (TSA)处理,细胞也被修饰。结果:结果表明,与CMV启动子相比,BLV启动子的基础活性较低,但在所有测试的细胞中,BLV启动子可以被诱导出与CMV启动子相似的高水平活性。Tax或BLV感染特异性增强BLVp活性,而对CMVp活性无影响。相比之下,非特异性激活剂TSA可以增强BLVp和CMVp的活性。结论:基于这些数据,我们认为BLV启动子在哺乳动物表达载体上的转基因表达是非常有用的。
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Comparison of bovine leukemia virus (BLV) and CMV promoter-driven reporter gene expression in BLV-infected and non-infected cells.

BACKGROUND: Viral promoters are used in mammalian expression vectors because they generally have strong activity in a wide variety of cells of differing tissues and species. METHODS: The utility of the BLV LTR/promoter (BLVp) for use in mammalian expression vectors was investigated through direct comparison to the CMV promoter (CMVp). Promoter activity was measured using luciferase assays of cell lines from different tissues and species stably transduced with BLVp or CMVp driven luciferase vectors including D17, FLK, BL3.1 and primary bovine B cells. Cells were also modified through the addition of BLV Tax expression vectors and/or BLV infection as well as treatment with trichostatin A (TSA). RESULTS: Results indicate the BLV promoter, while having low basal activity compared to the CMV promoter, can be induced to high-levels of activity similar to the CMV promoter in all cells tested. Tax or BLV infection specifically enhanced BLVp activity with no effect on CMVp activity. In contrast, the non-specific activator, TSA, enhanced both BLVp and CMVp activity. CONCLUSION: Based on these data, we conclude the BLV promoter could be very useful for transgene expression in mammalian expression vectors.

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