新城疫病毒HN蛋白膜外区的纯化

Purification of Envelope Domain of HN Protein from Newcastle Disease Virus Particles

血凝素神经氨酸酶(HN)是新城疫病毒(NDV)囊膜表面的糖蛋白之一,其在病毒表面的含量非常丰富,影响NDV的很多生物学特性,如热稳定性等。利用TritonX-100破坏病毒囊膜结构,分离囊膜糖蛋白,最后通过蛋白酶酶切和高速离心的方法获得了较为纯净的HN蛋白胞外区的蛋白样品,建立了从NDV粒子上纯化HN蛋白胞外区的方法。利用该方法获得的HN蛋白样品可以保持天然蛋白的结构、化学修饰和功能,能够更好地用于研究HN蛋白结构功能和生物学特性的形成机制,为新城疫病毒的基础研究、新城疫病毒载体改良应用和新城疫疫苗的研发奠定了基础。

Haemagglutinin-neuraminidase (HN) is one of two glycoproteins on the envelope surface of New-castle disease virus (NDV). The HN protein, which has very high abundance on the NDV surface, affects many biological characteristics of NDV, such as thermal stability and so on. In this paper,a method for purification of HN protein envelope domain from NDV particles was established;the NDV envelope was destroyed by TritonX-100 reagent and then envelope glycoproteins were isolated, finally,a relatively pure protein sample of HN protein extracellular domain was obtained by protease digestion and high speed centrifugation. The HN protein samples obtained by this method can maintain the structure , chemical modification and function of the natural protein,and can be better used to study the formation mechanism of the structure, function and biological characteristics of the HN protein, laying the foundation for the basic research of NDV ,the modified application of NDV vector and the development of NDV vaccine.