【摘要】 通过PCR的方法从猪基因组DNA中克隆IFN-β基因启动子片段,分别构建了含有猪β干扰素基因启动子萤光素酶报告质粒及其4个重复的NF-κB结合位点序列的萤光素酶报告质粒。脂质体基因转染法将萤光素酶报告质粒转染PK-15细胞,在poly(I∶C)或poly(dAT∶dAT)的刺激下,萤光素酶表达显著增加。本试验为进一步探讨猪β干扰素信号转导通路的研究奠定了基础。更多还原

【Abstract】 To establish a method for detection of porcine proteins and genes related to IFN-β signal transduction,we analyzed the regulatory elements that regulate the transcription of porcine IFN-β gene.The promoter region of porcine IFN-β gene and its four copies NF-κB binding site regions were amplified from porcine genomic DNA by PCR and were cloned into promoter-free plasmid pGL3 basic.Then these constructs were transiently transfected into PK-15 cells and luciferase activities were measured with or without the transfection of poly(I∶C) or poly(dAT∶dAT).Higher expression of luciferase was obviously detected in PK-15 cells transfected with poly(I∶C) or poly(dAT∶dAT).These reporter constructs are important tools for investigation of porcine IFN-β signaling transduction pathways.更多还原