Identification of pathogenic genes in very virulent herpesviruses by analysis on single-nucleotide polymorphism.

虽然有些疫苗正在使用，但应该开发新的更好的疫苗。在疫苗研制中，通过破坏毒株的毒力基因来构建无毒毒株是基本策略。但许多病毒的毒力基因尚未确定。毒力基因是根据毒株和非毒株之间的基因组多态性来确定的。在这项研究中，我们试图用单核苷酸多态性（snp）作为株间多态性标记来确定强毒疱疹病毒的毒力基因，这是人类基因组研究的热点。在寻找SNPs的过程中，病毒片段的制备被认为是最重要的一步，因为它适合于检测SNPs。虽然已经报道了许多检测snp的方法，但快速制备许多病毒DNA片段的方法尚未被报道为检测snp的一个步骤。因此，我们专注于开发制备多种类型病毒DNA片段的方法，并确定用于检测snp的片段的适当大小。最终得到如下结果，实现了本研究的第一个目的。首先，我们建立了一种独立于基因组序列的禽和哺乳动物疱疹病毒小瓶DNA片段的制备方法。其次，我们计算了疱疹病毒毒株和非毒株之间的snp，并确定了适合测定snp的大小。

Although some vaccines are in use, new superior vaccines should be developed. On the development of vaccines, construction of non-virulent strain by destruction of virulent genes of virulent strains is basic strategy. But virulent genes have not been determined in many virulent viruses. Virulent genes are determined by referring interstrain polymorphism of genome between virulent and non-virulent strains.In this study, we tried to determine virulent genes of very virulent herpesviruses using single-nucleotide polymorphisms (SNPs) as interstrain polymorphism markers, which is in the spotlight in human genomic research. On searching SNPs, preparation of viral fragments, which is suitable for detecting SNPs, is presumed to be most important step for detecting SNPs. Although many methods for detection of SNPs have been reported, method for rapid preparation of many viral DNA fragments have not been reported as a step for detection of SNPs. Thus, we focused on development of the method for preparation of many types of viral DNA fragments and also determination of appropriate size of the fragments to detect SNPs. Resultantly, we got the results below and achieved the first object of this research.Firstly, we developed a method for preparation of vial DNA fragments from avian and mammalian herpesviruses independent on its genomic sequences. Secondly, we counted SNPs between virulent and non-virulent strains of herpesviruses and determined the sizes appropriate for determination of SNPs.