目的：构建猪源SIRT1基因的过表达载体。方法：以GenBank中猪源SIRT1基因序列作为参考序列设计1对引物,通过IPEC-DQ细胞提取总RNA反转录成cDNA扩增SIRT1基因,构建pLVX-AcGFP1-N1-SIRT1克隆质粒,转染HEK 293T细胞并收集包装液,感染IPEC-DQ细胞,进行荧光检测和蛋白质免疫印迹检测。 结果：经PCR扩增、测序验证,pLVX-AcGFP1-N1-SIRT1重组表达质粒构建成功；荧光检测显示,感染含有pLVX-AcGFP1-N1-SIRT1重组质粒包装液的IPEC-DQ细胞中有明显绿色荧光信号。结论：成功构建猪源SIRT1基因的过表达载体,并在IPEC-DQ细胞中表达。

SIRT1基因；载体构建；IPEC-DQ细胞；基因表达

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江苏省优秀青年基金项目：“去乙酰化转移酶(SIRT1)调控猪流行性腹泻病毒逃逸先天免疫的作用机制”（BK20220114）。 国家自然科学基金项目：“沉默调节蛋白1调控猪流行性腹泻病毒逃逸先天免疫机制的研究”(32273015)。

DOI： https://doi.org/10.62639/sspis11.20250204

Published： 2025