Development of disease preventive method using radiated pathogenic microorganisms, cell lines and animals

The effects of radiation were investigated on pathogenic plasmid aiming at a development of a method to induce mutagenesis in plasmid DNA by radiation. To construct an experimental system which allows to detect a plasmid-segregated cell, kanamycin-resistant casette was inserted into pX02, a capsule plasmid in Bacillus anthracis to produce acpA:: Kmr by homologous recombination. This plasmid is thought available for analyzing the rate of plasmid segregation caused by radiation. Next, developments of detection and determination methods for various cytokines were attempted by RT-PCR method with an aim to investigate the expression changes of cytokine mRNA in calf immunocytes by radiation. In calf peripheral monocytes and alveolar macrophages, expressions of cytokine mRNAs such as IL-4, IFNα and GM-CSF mRNA as well as IL-1α, IL-1β, IL-2 and IL-6 were detected by RT-PCR method. (M.N.)