Possee R D, Howard S C
NERC Institute of Virology, Oxford, UK.
Nucleic Acids Res. 1987 Dec 23;15(24):10233-48. doi: 10.1093/nar/15.24.10233.
The polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus was analysed with respect to which sequences are required upstream of the mRNA transcription initiation (CAP) site for efficient promoter activity. Insertions (8, 95 and 785 nucleotides) were made in this region at an EcoR V site between the CAAT- and TATA-like boxes. When these mutations were introduced into the virus they did not affect the activity of the polyhedrin promoter as judged by expression of the beta-galactosidase (lacZ) gene inserted in lieu of the polyhedrin coding sequences. Deletions were made in the promoter which progressively removed sequences upstream from the CAP site. Removal of the TATA motif did not affect lacZ gene expression. A sequence 69 nucleotides upstream to the normal position of the polyhedrin ATG translation initiation codon was sufficient for maximum promoter activity but this was reduced by 90% when only 56 nucleotides upstream remained. The normal CAP site was utilized by each deletion mutant. Promoter activity was undetectable when the CAP site was deleted. The results are discussed in relation to other eukaryotic promoters.
针对加利福尼亚苜蓿银纹夜蛾核型多角体病毒的多角体蛋白基因启动子，分析了在mRNA转录起始（CAP）位点上游哪些序列对于高效启动子活性是必需的。在该区域位于类CAAT盒和类TATA盒之间的EcoR V位点进行了插入（8、95和785个核苷酸）。当将这些突变引入病毒时，根据代替多角体蛋白编码序列插入的β-半乳糖苷酶（lacZ）基因的表达判断，它们并未影响多角体蛋白启动子的活性。在启动子中进行了缺失操作，逐步去除CAP位点上游的序列。去除TATA基序并不影响lacZ基因表达。多角体蛋白ATG翻译起始密码子正常位置上游69个核苷酸的序列足以实现最大启动子活性，但当仅保留上游56个核苷酸时，活性降低了90%。每个缺失突变体都利用了正常的CAP位点。当CAP位点缺失时，无法检测到启动子活性。结合其他真核生物启动子对结果进行了讨论。
Zotero 插件
