摘要
目的:构建天然免疫胞内识别受体核苷酸寡聚域1(NOD1)真核表达质粒。方法:NOD1基因片段经PCR扩增获得,经酶切后连接到真核表达载体pcDNA3/flag中,对挑选出的阳性克隆测序,将序列正确的重组质粒pflag-NOD1转染293T细胞,用Western印迹检测目的蛋白的表达,同时用NF-κB的萤光素酶报告基因检测NOD1蛋白的活性。结果:pflag-NOD1可以在真核细胞293T中表达,并可以增强NF-κB报告基因的转录活性。结论:构建了重组质粒pflag-NOD1,在细胞中表达NOD1后能够提高NF-κB转录的生物活性,为进一步研究NOD1的功能奠定了基础。
Objective：To construct the eukaryotic expression plasmid of nucleotide oligomerization domain1 （NOD1）.Methods：NOD1 gene fragment was amplified by PCR,and was ligated into pcDNA3 /flag vector.The transformed clones were sequenced and the right one named pflag-NOD1 was transfected into 293T cell line to test the expression of NOD1,and the activity of NOD1 by Western blot and luciferase analysis.Results：pflagNOD1 expressed NOD1 in 293T cell line,and induced high activity of NF-κB transcription.Conclusion：Recombinant plasmid pflag-NOD1 was constructed successfully and the NOD1 expressed had the activity of inducing NF-κB transcription.
出处
《生物技术通讯》
CAS
2010年第2期184-187,共4页
Letters in Biotechnology
关键词
NOD1
NF-ΚB
真核表达
萤光素酶报告基因
nucleotide oligomerization domain 1 NF-κB eukaryotic expression luciferase reporter gene
