Toyoda T, Wolffe A P
Laboratory of Molecular Embryology, National Institutes of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Dev Biol. 1992 Sep;153(1):150-7. doi: 10.1016/0012-1606(92)90099-3.
We examine the RNA polymerase II-dependent transcription directed by several promoters in extracts prepared from distinct developmental stages of Xenopus laevis. RNA polymerase II accurately initiates transcription from the cytomegalovirus, herpes simplex virus thymidine kinase, and Xenopus heat-shock protein (hsp) 70 promoters. The efficiency of transcription of these different promoters is dependent on whether extracts from oocytes, eggs, or somatic cells are used and on the temperature of incubation. In contrast to the viral promoters, the hsp 70 promoter is more active at heat shock temperatures in oocyte and egg extracts (31 degrees-34 degrees C) than at physiological temperatures for Xenopus (20 degrees-25 degrees C). These in vitro transcription extracts should be useful in examining the molecular mechanisms responsible for differential gene expression during Xenopus development.
我们在从非洲爪蟾不同发育阶段制备的提取物中,研究了由几种启动子指导的RNA聚合酶II依赖性转录。RNA聚合酶II能准确地从巨细胞病毒、单纯疱疹病毒胸苷激酶和非洲爪蟾热休克蛋白(hsp)70启动子起始转录。这些不同启动子的转录效率取决于所使用的是卵母细胞、卵子还是体细胞的提取物,以及孵育温度。与病毒启动子不同,hsp 70启动子在卵母细胞和卵子提取物的热休克温度(31摄氏度 - 34摄氏度)下比在非洲爪蟾的生理温度(20摄氏度 - 25摄氏度)下更具活性。这些体外转录提取物将有助于研究非洲爪蟾发育过程中基因差异表达的分子机制。