Patel G, Jones N C
Gene Regulation Laboratory, Imperial Cancer Research Fund, London, UK.
Nucleic Acids Res. 1990 May 25;18(10):2909-15. doi: 10.1093/nar/18.10.2909.
The baculovirus expression system has been successfully used to overproduce a number of different protein products. In this report we describe the construction of a recombinant baculovirus containing the adenovirus E1A 13s cDNA sequence. Infection of insect cells with this virus results in the production of phosphorylated E1A protein. The phosphorylation pattern appears to be similar to the complex pattern associated with E1A protein synthesis in mammalian cells. Purified baculovirus generated E1A protein activated transcription of specific poIIII promoters both in microinjected Xenopus laevis oocytes and in HeLa cell in vitro transcription extracts. The protein also stimulates in vitro transcription of the poIIII transcribed VA1 gene.
杆状病毒表达系统已成功用于大量生产多种不同的蛋白质产物。在本报告中,我们描述了一种含有腺病毒E1A 13s cDNA序列的重组杆状病毒的构建。用这种病毒感染昆虫细胞会导致磷酸化E1A蛋白的产生。磷酸化模式似乎与哺乳动物细胞中与E1A蛋白合成相关的复杂模式相似。纯化的杆状病毒产生的E1A蛋白在显微注射的非洲爪蟾卵母细胞和HeLa细胞体外转录提取物中均激活了特定的RNA聚合酶III启动子的转录。该蛋白还刺激了RNA聚合酶III转录的VA1基因的体外转录。