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首页> 外文期刊>Chinese science bulletin >Three-dimensional reconstruction of mature dengue virus cultured in neutral medium
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Three-dimensional reconstruction of mature dengue virus cultured in neutral medium

机译：在中性培养基中培养的成熟登革热病毒的三维重建

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Dengue virus (DEN), a single positively stranded RNA virus, is a family member of Flaviviridae; it uses the mosquito Aedes aegypti as its principle vector to cause what is known as Dengue fever. Its genome is approx 10.8-kb in size and has one open reading frame encoding three structural proteins: capsid (C), precursor of the membrane (prM) and envelope (E), as well as seven non-structural proteins from a single polypeptide. The immature DEN particles are assembled on the inner surface of the endoplasmic reticulum(ER) and budded into the lumen of the ER. The diameter of immature particle is approximately 60 nm and the envelope is composed of 60 trimers of prM-E heterodimers. The prM proteins are cleaved into M proteins by furin during their transportation through the trans-Golgi network (TGN), resulting in changes of both the conformation and arrangement of envelope proteins and yielding mature virus particles, the surface of which is compactly lined up with 90 E-E homodimers. The diameter of the mature virus shrinks to 50 nm. In order to get the virus particles of relatively homogeneous configuration, various methods have been employed during the preparation of virus culture. The addition of furin in acid culture can stimulate the yield of mature virus particles while adding NH_4Cl improves the yield of immature virus particles due to the inhibited cleavage of prM by furin. In this work, the DEN viruses were cultured and purified under neutral conditions without further addition of furin or NH_4Cl in order to study the possible conformational changes of DEN II.
机译：登革热病毒（DEN）是一种正链RNA病毒，是黄病毒科的一个家族成员。它以蚊子埃及伊蚊为主要媒介引起登革热。它的基因组大小约为10.8-kb，具有一个开放阅读框，其编码三种结构蛋白：衣壳（C），膜前体（prM）和包膜（E），以及来自单个多肽的7种非结构蛋白。未成熟的DEN粒子聚集在内质网（ER）的内表面上，并萌芽到ER的管腔中。未成熟颗粒的直径约为60 nm，包膜由60个三聚体prM-E异二聚体组成。 prM蛋白在通过反式高尔基体网络（TGN）转运过程中被弗林蛋白酶切割成M蛋白，导致包膜蛋白的构象和排列发生变化，并产生成熟的病毒颗粒，其表面紧密排列90 EE同二聚体。成熟病毒的直径缩小到50 nm。为了获得相对均一构型的病毒颗粒，在病毒培养物的制备过程中已经采用了各种方法。在酸性培养物中添加弗林蛋白酶可以刺激成熟病毒颗粒的产量，而由于弗林蛋白酶抑制prM的裂解，添加NH_4Cl可以提高未成熟病毒颗粒的产量。在这项工作中，在不进一步添加弗林蛋白酶或NH_4Cl的情况下，在中性条件下培养和纯化DEN病毒，以研究DEN II可能的构象变化。

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