鱼类弹状病毒的NV基因：用于快速评估遗传变异的核糖核酸酶保护分析方法的开发

The NV genes of fish rhabdoviruses: development of RNase protection assays for rapid assessment of genetic variation.

作者信息

Kurath G, Higman K H, Björklund H V

机构信息

National Biological Service, Northwest Biological Science Center, Seattle, WA 98115, USA.

出版信息

Vet Res. 1995;26(5-6):477-85.

PMID:8581026

Abstract

The genomic RNA of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV) contains a small NV gene which is not present in any other rhabdoviruses characterized to date. We have constructed a plasmid which carries a full length cDNA copy of the IHNV NV gene between 2 RNA polymerase promoters such that plus-sense and minus-sense RNA transcripts of the IHNV NV gene can be synthesized. These were used to develop an RNase protection assay which was capable of detecting as little as 1-5 ng of NV mRNA or genomic RNA. RNA from tissue culture cells infected with 3 different strains of IHNV showed distinct, diagnostic cleavage patterns reflecting sequence variation in their NV genes.

摘要

鱼类弹状病毒传染性造血器官坏死病毒（IHNV）的基因组RNA含有一个小的NV基因，该基因在迄今已鉴定的任何其他弹状病毒中均不存在。我们构建了一个质粒，该质粒在两个RNA聚合酶启动子之间携带IHNV NV基因的全长cDNA拷贝，从而可以合成IHNV NV基因的正义和反义RNA转录本。这些被用于开发一种核糖核酸酶保护分析方法，该方法能够检测低至1-5 ng的NV mRNA或基因组RNA。来自感染了3种不同IHNV毒株的组织培养细胞的RNA显示出独特的、具有诊断意义的切割模式，反映了其NV基因的序列变异。