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新城疫病毒基质蛋白的异源表达、鉴定及作为抗病毒治疗靶点的评估。

Heterologous expression, characterization and evaluation of the matrix protein from Newcastle disease virus as a target for antiviral therapies.

出版信息

Appl Microbiol Biotechnol. 2014 Feb;98(4):1691-701. doi: 10.1007/s00253-013-5043-2. Epub 2013 Jun 25.

Abstract

Newcastle disease virus (NDV) is an infectious agent of a large variety of birds, including chicken, which poses a real threat to the agriculture industry. Matrix (M) proteins of NDV and many other viruses perform critical functions during viral assembly and budding from the host cell. M-proteins are well conserved and therefore are potential targets for antiviral therapies. To validate this, we expressed the NDV M-protein in its native form in Saccharomyces cerevisiae and in inclusion bodies in Escherichia coli. Proper refolding of the recombinant protein produced in E. coli was verified using circular dichroism and infrared spectroscopies and electron microscopy. Immunization of chickens with the NDV M-protein elicited significant serum antibody titers. However, the antibodies conferred little protection against the ND following lethal viral challenges. We conclude that the M-protein is not exposed on the surface of the host cell or the virus at any stage during its life cycle. We discuss how the conserved M-protein can further be exploited as an antiviral drug target.

摘要

新城疫病毒(NDV)是一种感染多种鸟类的病原体,包括鸡,这对农业产业构成了真正的威胁。NDV 和许多其他病毒的基质(M)蛋白在病毒组装和从宿主细胞出芽过程中发挥关键作用。M 蛋白高度保守,因此是抗病毒治疗的潜在靶点。为了验证这一点,我们在酿酒酵母中以天然形式和在大肠杆菌中以包涵体形式表达了 NDV M 蛋白。使用圆二色性和红外光谱学以及电子显微镜验证了在大肠杆菌中产生的重组蛋白的正确重折叠。用 NDV M 蛋白免疫鸡可引起显著的血清抗体滴度。然而,这些抗体在致命病毒攻击后对 ND 的保护作用很小。我们得出的结论是,在其生命周期的任何阶段,M 蛋白都不会暴露在宿主细胞或病毒的表面。我们讨论了如何进一步利用保守的 M 蛋白作为抗病毒药物靶点。

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