猪NLRP3基因克隆及其在DF-1细胞的表达
发布时间：2018-11-15 09:43
【摘要】：NLRP3蛋白作为NLRP3炎性小体的主要组成蛋白之一,在机体对抗病原微生物和炎症反应中起着重要作用。为了深入研究猪的NLRP3蛋白在炎性疾病发生过程中的作用机制,采用PCR的方法扩增了长白猪NLRP3基因,并对其序列和编码蛋白的结构进行了分析。构建了pIRES2-EGFP-NLRP3真核表达载体,并转染至DF-1细胞中进行表达。结果显示,该基因表现出种属内保守性和种属间差异性,编码蛋白由1 036个氨基酸构成,存在4个潜在跨膜区,分别是第401~419,第721~739,第775~795和第889~910位氨基酸,不含信号肽。单层细胞免疫化学检测结果显示,重组质粒pIRES2-EGFP-NLRP3成功转染并在细胞中表达。本研究为深入探讨炎症小体在猪炎症性疾病的作用机制提供了基础数据。
[Abstract]:As one of the major components of NLRP3 inflammatory bodies, NLRP3 protein plays an important role in the fight against pathogenic microorganisms and inflammation. In order to study the role of NLRP3 protein in the pathogenesis of inflammatory diseases in pigs, the NLRP3 gene of Landrace pig was amplified by PCR, and its sequence and protein structure were analyzed. PIRES2-EGFP-NLRP3 eukaryotic expression vector was constructed and transfected into DF-1 cells for expression. The results showed that the gene showed intra-species conservation and inter-species difference. The encoded protein was composed of 1,036 amino acids and had four potential transmembrane regions, namely, the 401r-419, 721-, 739-, 775-, 889-, and -910 amino acids, which contained no signal peptide. The results of monolayer immunocytochemistry showed that the recombinant plasmid pIRES2-EGFP-NLRP3 was successfully transfected and expressed in the cells. This study provides basic data for further study of the mechanism of inflammatory bodies in porcine inflammatory diseases.
【作者单位】： 华南农业大学兽医学院;
【基金】：农业部公益性行业资助项目(201203039)
【分类号】：S858.28
，
本文编号：2332938
[Abstract]:As one of the major components of NLRP3 inflammatory bodies, NLRP3 protein plays an important role in the fight against pathogenic microorganisms and inflammation. In order to study the role of NLRP3 protein in the pathogenesis of inflammatory diseases in pigs, the NLRP3 gene of Landrace pig was amplified by PCR, and its sequence and protein structure were analyzed. PIRES2-EGFP-NLRP3 eukaryotic expression vector was constructed and transfected into DF-1 cells for expression. The results showed that the gene showed intra-species conservation and inter-species difference. The encoded protein was composed of 1,036 amino acids and had four potential transmembrane regions, namely, the 401r-419, 721-, 739-, 775-, 889-, and -910 amino acids, which contained no signal peptide. The results of monolayer immunocytochemistry showed that the recombinant plasmid pIRES2-EGFP-NLRP3 was successfully transfected and expressed in the cells. This study provides basic data for further study of the mechanism of inflammatory bodies in porcine inflammatory diseases.
【作者单位】： 华南农业大学兽医学院;
【基金】：农业部公益性行业资助项目(201203039)
【分类号】：S858.28
，
本文编号：2332938
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