湖北钉螺髓样分化因子88的鉴定及其在抗血吸虫感染固有免疫中的地位
发布时间：2019-06-18 11:41
【摘要】：目的克隆、鉴定湖北钉螺(Oncomelania hupensis)髓样分化因子88(MyD88)基因,并通过观察感染日本血吸虫前后钉螺各组织中MyD88 m RNA表达水平的变化,探讨其在抗血吸虫感染固有免疫中的地位。方法通过c DNA末端快速扩增技术(Rapid amplification of c DNA ends,RACE)获取湖北钉螺MyD88全长c DNA序列,预测其蛋白结构域,并进行多序列比对及保守区域分析,构建系统进化树。使用实时荧光定量PCR(Real-time quantitative PCR,RT-q PCR)技术检测MyD88基因在血吸虫感染前后钉螺各组织中的表达变化。结果湖北钉螺MyD88全长c DNA开放阅读框(Open reading frame,ORF)1 406 bp,编码468个氨基酸,蛋白N端和C端分别存在死亡结构域和TIR(Toll/interlrukin-1 receptor,TIR)结构域。与其他软体类MyD88氨基酸序列比对,相似性为38%~52%;系统进化树提示钉螺MyD88和光滑双脐螺MyD88起源于共同的祖先基因。q PCR结果显示,检测的所有组织中均有MyD88 m RNA的表达,其中血淋巴细胞中表达最为丰富。感染日本血吸虫后,除钉螺头足外,MyD88 m RNA在肝脏、生殖腺、血淋巴细胞等组织中均有上调表达,以血淋巴细胞中上调表达最显著。结论湖北钉螺存在依赖MyD88的TLRs(Toll-like receptors,TLRs)信号通路,MyD88分子可能在钉螺对抗日本血吸虫感染的固有免疫中发挥重要作用。
[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，
本文编号：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，
本文编号：2501460
本文链接：https://www.wllwen.com/xiyixuelunwen/2501460.html
最近更新
教材专著
