湖北釘螺髓樣分化因子88的鑒定及其在抗血吸蟲感染固有免疫中的地位
發(fā)布時間：2019-06-18 11:41
【摘要】：目的克隆、鑒定湖北釘螺(Oncomelania hupensis)髓樣分化因子88(MyD88)基因,并通過觀察感染日本血吸蟲前后釘螺各組織中MyD88 m RNA表達(dá)水平的變化,探討其在抗血吸蟲感染固有免疫中的地位。方法通過c DNA末端快速擴(kuò)增技術(shù)(Rapid amplification of c DNA ends,RACE)獲取湖北釘螺MyD88全長c DNA序列,預(yù)測其蛋白結(jié)構(gòu)域,并進(jìn)行多序列比對及保守區(qū)域分析,構(gòu)建系統(tǒng)進(jìn)化樹。使用實時熒光定量PCR(Real-time quantitative PCR,RT-q PCR)技術(shù)檢測MyD88基因在血吸蟲感染前后釘螺各組織中的表達(dá)變化。結(jié)果湖北釘螺MyD88全長c DNA開放閱讀框(Open reading frame,ORF)1 406 bp,編碼468個氨基酸,蛋白N端和C端分別存在死亡結(jié)構(gòu)域和TIR(Toll/interlrukin-1 receptor,TIR)結(jié)構(gòu)域。與其他軟體類MyD88氨基酸序列比對,相似性為38%~52%;系統(tǒng)進(jìn)化樹提示釘螺MyD88和光滑雙臍螺MyD88起源于共同的祖先基因。q PCR結(jié)果顯示,檢測的所有組織中均有MyD88 m RNA的表達(dá),其中血淋巴細(xì)胞中表達(dá)最為豐富。感染日本血吸蟲后,除釘螺頭足外,MyD88 m RNA在肝臟、生殖腺、血淋巴細(xì)胞等組織中均有上調(diào)表達(dá),以血淋巴細(xì)胞中上調(diào)表達(dá)最顯著。結(jié)論湖北釘螺存在依賴MyD88的TLRs(Toll-like receptors,TLRs)信號通路,MyD88分子可能在釘螺對抗日本血吸蟲感染的固有免疫中發(fā)揮重要作用。
[Abstract]:Objective to clone and identify the (Oncomelania hupensis) myeloid differentiation factor 88 (MyD88) gene of Oncomelania hupensis (Oncomelania hupensis), and to observe the expression of MyD88 m RNA in the tissues of Oncomelania hupensis before and after infection with Schistosoma Japonicum, and to explore its role in the innate immunity against Schistosoma japonicum infection. Methods the full-length c-DNA sequence of Oncomelania hupensis MyD88 was obtained by c-DNA terminal rapid amplification (Rapid amplification of c DNA ends,RACE). The protein domain was predicted, and the multiple sequence alignment and conservative region analysis were carried out to construct the phylogenetic tree. The expression of MyD88 gene in snail tissues before and after schistosomiasis infection was detected by real-time fluorescence quantitative PCR (Real-time quantitative PCR,RT-q PCR). Results the full length c DNA open reading frame (Open reading frame,ORF) 1.406 bp, of Oncomelania hupensis MyD88 encodes 468 amino acids. The N terminal and C terminal of the protein have death domain and TIR (Toll/interlrukin-1 receptor,TIR) domain, respectively. Compared with other soft MyD88 amino acid sequences, the similarity was 38% 鈮，
本文編號：2501460
[Abstract]:Objective to clone and identify the (Oncomelania hupensis) myeloid differentiation factor 88 (MyD88) gene of Oncomelania hupensis (Oncomelania hupensis), and to observe the expression of MyD88 m RNA in the tissues of Oncomelania hupensis before and after infection with Schistosoma Japonicum, and to explore its role in the innate immunity against Schistosoma japonicum infection. Methods the full-length c-DNA sequence of Oncomelania hupensis MyD88 was obtained by c-DNA terminal rapid amplification (Rapid amplification of c DNA ends,RACE). The protein domain was predicted, and the multiple sequence alignment and conservative region analysis were carried out to construct the phylogenetic tree. The expression of MyD88 gene in snail tissues before and after schistosomiasis infection was detected by real-time fluorescence quantitative PCR (Real-time quantitative PCR,RT-q PCR). Results the full length c DNA open reading frame (Open reading frame,ORF) 1.406 bp, of Oncomelania hupensis MyD88 encodes 468 amino acids. The N terminal and C terminal of the protein have death domain and TIR (Toll/interlrukin-1 receptor,TIR) domain, respectively. Compared with other soft MyD88 amino acid sequences, the similarity was 38% 鈮，
本文編號：2501460
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