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经典的Wnt信号通路是一条与生长发育密切相关的信号通路,经典Wnt信号通路在棉铃虫滞育蛹脑中受到抑制。本研究以棉铃虫Wnt1基因的转录调控为研究目标,对Wnt1启动子的活性进行了分析。首先,运用染色体步移的技术成功克隆了一段长为1566 bp的Wnt1启动子。然后,通过PCR扩增不同长度的启动子片段,构建到报告质粒,转染棉铃虫Hz Am1细胞并进行启动子活性分析,结果表明-1077~-1120以及-1120~-1140这两个区域对Wnt1基因的转录起着明显的激活作用。最后对这两个区域潜在的转录因子结合位点进行了预测。本研究从根本上了解棉铃虫滞育蛹脑中经典Wnt信号通路的调节机制打下了基础。
The classical Wnt signaling pathway is a closely related signal pathway to growth and development. The classical Wnt signaling pathway is inhibited in the diapause brain of cotton bollworm. In this study, we investigated the transcriptional regulation of Wnt1 gene in Helicoverpa armigera and analyzed the activity of Wnt1 promoter. First, a 1566 bp Wnt1 promoter was successfully cloned using chromosome walking technology. Then, different lengths of promoter fragments were amplified by PCR, constructed into reporter plasmids, and transfected into Helicoverpa armigera Hz Am1 cells and analyzed for promoter activity. The results showed that -1077 ~ -1120 and -1120 ~ -1140 Wnt1 gene transcription plays a significant role in activation. Finally, the potential transcription factor binding sites in these two regions were predicted. This study laid the foundation for a fundamental understanding of the regulatory mechanism of the canonical Wnt signaling pathway in the diapause pupae of H. armigera.