【摘要】：NLRP3蛋白作為NLRP3炎性小體的主要組成蛋白之一,在機(jī)體對(duì)抗病原微生物和炎癥反應(yīng)中起著重要作用。為了深入研究豬的NLRP3蛋白在炎性疾病發(fā)生過(guò)程中的作用機(jī)制,采用PCR的方法擴(kuò)增了長(zhǎng)白豬NLRP3基因,并對(duì)其序列和編碼蛋白的結(jié)構(gòu)進(jìn)行了分析。構(gòu)建了pIRES2-EGFP-NLRP3真核表達(dá)載體,并轉(zhuǎn)染至DF-1細(xì)胞中進(jìn)行表達(dá)。結(jié)果顯示,該基因表現(xiàn)出種屬內(nèi)保守性和種屬間差異性,編碼蛋白由1 036個(gè)氨基酸構(gòu)成,存在4個(gè)潛在跨膜區(qū),分別是第401~419,第721~739,第775~795和第889~910位氨基酸,不含信號(hào)肽。單層細(xì)胞免疫化學(xué)檢測(cè)結(jié)果顯示,重組質(zhì)粒pIRES2-EGFP-NLRP3成功轉(zhuǎn)染并在細(xì)胞中表達(dá)。本研究為深入探討炎癥小體在豬炎癥性疾病的作用機(jī)制提供了基礎(chǔ)數(shù)據(jù)。
[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.
【作者單位】： 華南農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院;
【基金】：農(nóng)業(yè)部公益性行業(yè)資助項(xiàng)目(201203039)
【分類號(hào)】：S858.28
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本文編號(hào)：2332938