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Sequence Analysis of the NS Gene of an NS Mutant of Influenza B Virus.

乙型流感病毒 NS 突变体的 NS 基因的序列分析。

基本信息

批准号：
59480173
负责人：
TOBITA Kiyotake
金额：
$ 3.71万
依托单位：
Jichi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59480173/
关键词：
Influenza B Virus Viral Protein Viral RNA NS Gene Sequencing 欠落変異

项目摘要

By repeated back-crosses of influenza virus A/Aichi/2/68 with wild type B/Yamagata/1/73, we isolated a B type virus mutant, clone 201, bearing mutations in the NS gene ( RNA segment 8 ). Clone 201 grew efficiently both in MDCK cells and in fertile hens' eggs, and induced a severe cytopathic effect in MDCK cells early after infection, which was ascribed to the function of the mutated NS gene of clone 201 by reassortment experiment.Sequencing analysis of the NS gene of clone 201 revealed that there is a deletion of 13 bases from position 374 - 386 of the plus sense RNA, and, due to the frame shift downstream, the translation of <NS_1> stops at position 425, giving rise to an <NS_1> polypeptide consisting of 127 amino acids, more than a half shorter than the <NS_1> of wild type B/Yamagata ( 281 amino acids ). In addition to the deletion, 10 point mutations were noticed, including 4 silent mutations.The sequencing data are consisting with the findings that RNA segment 8 of 201 migrates slightly faster than the wild type counterpart in the polyacrylamide gel, and that 201 lacks the protein band for the <NS_1> of B/Yamagata ( Molecular Weight 39,800 ) and a heavy band is present instead at the position of <NS_2> of B/Yamagata ( Molecular Weight 19,000 ).Whether severe cytolysis induced by clone 201 is due to the large carboxy terminal deletion of <NS_1> or to the point mutation(s) is yet to be determined.

通过流感病毒 A/Aichi/2/68 与野生型 B/Yamagata/1/73 的重复回交，我们分离出 B 型病毒突变体，克隆 201，其 NS 基因（RNA 片段 8）发生突变。克隆201在MDCK细胞和受精鸡蛋中均能高效生长，并在感染后早期在MDCK细胞中诱导严重的细胞病变效应，通过重配实验将其归因于克隆201突变的NS基因的功能。对克隆201的NS基因的测序分析显示，在正义的374-386位点存在13个碱基的缺失。 RNA，并且由于移码下游，<NS_1>的翻译在位置425处停止，产生由127个氨基酸组成的<NS_1>多肽，比野生型B/Yamagata的<NS_1>（281个氨基酸）短一半以上。除了缺失之外，还发现了 10 个点突变，其中包括 4 个沉默突变。测序数据显示，201 的 RNA 片段 8 在聚丙烯酰胺凝胶中的迁移速度略快于野生型对应物，并且 201 缺乏 B/Yamagata 的 <NS_1>（分子量 39,800）的蛋白质条带，而在 B/Yamagata 的 <NS_2> 位置存在重条带。 B/Yamagata（分子量19,000）。克隆201引起的严重细胞溶解是否是由于<NS_1>的羧基末端大缺失或点突变引起的，尚待确定。