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

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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.