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Cloning expression and functional analysis of the duck Toll-like receptor 5 (TLR5) gene

机译:鸭Toll样受体5(TLR5)基因的克隆表达及功能分析

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摘要

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In the present study, the first TLR5 gene in duck was cloned. The open reading frame (ORF) of duck TLR5 (dTLR5) cDNA is 2580 bp and encodes a polypeptide of 859 amino acids. We also cloned partial sequences of myeloid differentiation factor 88, 2'-5'-oligoadenylate synthetase (OAS), and myxovirus resistance (Mx) genes from duck. dTLR5 mRNA was highly expressed in the bursa of Fabricius, spleen, trachea, lung, jejunum, rectum, and skin; moderately expressed in the muscular and glandular tissues, duodenum, ileum, caecum, and pancreas; and minimally expressed in the heart, liver, kidney, and muscle. DF-1 or HeLa cells transfected with DNA constructs encoding dTLR5 can activate NF-κB leading to the activation of interleukin-6 (IL-6) promoter. When we challenged ducks with a Herts33 Newcastle disease virus (NDV), mRNA transcription of the antiviral molecules Mx, Double stranded RNA activated protein kinase (PKR), and OAS was up-regulated in the liver, lung, and spleen 1 and 2 days post-inoculation.
机译:Toll样受体5(TLR5)负责识别脊椎动物中细菌鞭毛蛋白。在本研究中,克隆了鸭中的第一个TLR5基因。鸭TLR5(dTLR5)cDNA的开放阅读框(ORF)为2580 bp,编码859个氨基酸的多肽。我们还克隆了鸭的髓系分化因子88、2'-5'-寡腺苷酸合成酶(OAS)和粘液病毒抗性(Mx)基因的部分序列。 dTLR5 mRNA在法氏囊,脾,气管,肺,空肠,直肠和皮肤的法氏囊中高表达。在肌肉和腺体组织,十二指肠,回肠,盲肠和胰腺中适度表达;并在心脏,肝脏,肾脏和肌肉中很少表达。用编码dTLR5的DNA构建体转染的DF-1或HeLa细胞可以激活NF-κB,从而激活白介素6(IL-6)启动子。当我们用Herts33新城疫病毒(NDV)攻击鸭子时,抗病毒分子Mx,双链RNA活化蛋白激酶(PKR)和OAS的mRNA转录在肝,肺和脾中分别上调了1天和2天接种后。

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