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首页> 外文期刊>Journal of Molecular Biology >Construction and crystal structure of recombinant STNV capsids.
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Construction and crystal structure of recombinant STNV capsids.

机译：重组STNV衣壳的构建和晶体结构。

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摘要

A codon-optimised gene has been expressed in Escherichia coli to produce the coat protein (CP) of the Satellite Tobacco Necrosis Virus. This protein assembles in vivo into capsids closely resembling those of the T=1 wild-type virus. These virus-like particles (VLPs) package the recombinant mRNA transcript and can be disassembled and reassembled using different buffer conditions. The X-ray crystal structure of the VLP has been solved and refined at 1.4 A resolution and shown to be very similar to that of wild-type Satellite Tobacco Necrosis Virus, except that icosahedral symmetry constraints could be removed to reveal differences between subunits, presumably owing to crystal packing. An additional low-resolution X-ray crystal structure determination revealed well-ordered RNA fragments lodged near the inside surface of the capsid, close to basic clusters formed by the N-terminal helices that project into the interior of the particle. The RNA consists of multiple copies of a 3-bp helical stem, with a single unpaired base at the 3' end, and probably consists of a number of short stem-loops where the loop region is disordered. The arrangement of the RNA is different from that observed in other satellite viruses.
机译：密码子优化的基因已在大肠杆菌中表达，以产生卫星烟草坏死病毒的外壳蛋白（CP）。该蛋白质在体内组装成衣壳，其衣壳与T = 1野生型病毒的衣壳极为相似。这些病毒样颗粒（VLP）包装了重组mRNA转录物，可以使用不同的缓冲液条件进行拆卸和重新组装。解决了VLP的X射线晶体结构，并以1.4 A的分辨率对其进行了改进，并显示出与野生型卫星烟草坏死病毒非常相似，不同之处在于可以消除二十面体对称性约束以揭示亚基之间的差异由于晶体包装。另外的低分辨率X射线晶体结构测定显示，衣壳内部表面附近排列有序的RNA片段，靠近由伸入颗粒内部的N末端螺旋形成的基本簇。 RNA由3 bp螺旋茎的多个副本组成，在3'端具有一个不成对的碱基，并且可能由许多短茎环组成，其中环区无序。 RNA的排列不同于其他卫星病毒中观察到的排列。

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