Basurco B, Benmansour A
Unité de Virologie et Immunologie Moléculaires, INRA, Jouy-En-Josas, France.
Virology. 1995 Oct 1;212(2):741-5. doi: 10.1006/viro.1995.1534.
We used direct RNA sequencing to determine the genomic organization of the region downstream from the G gene of viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus. This region contains a gene coding for a protein, identified as nonvirion protein (NV), and the gene coding for the RNA polymerase (L). Thus, VHSV genome organization was confirmed to be 3'-N-P-M-G-NV-L-5'. In both a virulent European (07-71) and an avirulent North American (Makah) strain, the NV gene is transcribed into a small mRNA that codes for a protein of 122 amino acids. It has no significant sequence similarity with the infectious hematopoietic necrosis virus NV protein nor with any other known protein. We expressed the NV protein as a fusion protein with the glutathione S-transferase of Schistosoma japonicum and used the purified fusion protein to immunize rabbits. The rabbit antiserum precipitated from infected cell extracts--and not from noninfected cells or purified virions--a protein of 14 kDa, well in accordance with the expected NV gene product size. The prediction that the NV protein is a nonstructural protein is supported by its absence from mature virions although it is present in infected cells.
我们使用直接RNA测序来确定病毒性出血性败血症病毒（VHSV，一种鱼类弹状病毒）G基因下游区域的基因组结构。该区域包含一个编码蛋白质的基因，被鉴定为非病毒粒子蛋白（NV），以及编码RNA聚合酶（L）的基因。因此，VHSV的基因组结构被确认为3'-N-P-M-G-NV-L-5'。在一个强毒力的欧洲毒株（07-71）和一个无毒力的北美毒株（马卡）中，NV基因都转录成一种小mRNA，其编码一个由122个氨基酸组成的蛋白质。它与传染性造血坏死病毒的NV蛋白以及任何其他已知蛋白均无显著的序列相似性。我们将NV蛋白表达为与日本血吸虫谷胱甘肽S-转移酶的融合蛋白，并使用纯化的融合蛋白免疫兔子。兔抗血清能从感染细胞提取物中沉淀出一种14 kDa的蛋白质，而不能从未感染细胞或纯化病毒粒子中沉淀出该蛋白，这与预期的NV基因产物大小完全相符。尽管NV蛋白存在于感染细胞中，但在成熟病毒粒子中不存在，这一事实支持了NV蛋白是非结构蛋白的预测。
