KOJIMA Kazunobu Sapporo Med.Univ., Dept.of Hygiene, Instructor, 医学部, 助手 (20264517)
KOBAYASHI Nobumichi Sapporo Med.Univ., Dept.of Hygiene, Associate Professor, 医学部, 助教授 (80186759)
URASAWA Tomoko Sapporo Med.Univ., Dept.of Hygiene, Professor, 保健医療学部, 教授 (90045378)
|Budget Amount *help
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥3,900,000 (Direct Cost: ¥3,900,000)
1.We isolaed subclones producing large, medium, or small plaques from each of A5-13 expressing normal NSP1, A5-10 encoding omly 40 amino acid-NSP1, and A5-16 having NSP1 gene with 500 bp deletion. All of the clones replicated well in MA-104 cells, and induced dirrhea in suckling mice by oral administration. These results indicate that the insertion of foreign gene into NSP1 gene will be possible when the system of artificial infectious virus particles is established.
2.We first determined the complete nucleotide sequence of the genome of human ratavirus (strain KU).
3.All of the 6 structural proteins VP1-VP4, VP6, VP7 and 4 of 5 non-structural proteins of human rotavirus were expressed in baculovirus expression system. The produre for purification of his-tagged nonstructural proteins was established, and this enabled the precise examination of the function of nonstructural proteins.
4.Self-assembled, artificial and empty single-shelled particles or double-shelled particules were obtained by coexpressions of VP2 and VP6, or VP2, VP6 and VP7. In addition, the coexpression of VP1, VP3, VP6, VP7, and VP4 as well as VP2 resulted in the formation of artificial virus-like particles lacking genome. These artificial empty particles have the value as a value as a vaccine candidate and a tool for studying the immunity of rotavirus infection.
5.In the infection experiments by oral inoculation to suckling mice, we observed the diarrhea in the mice administered with G1, G3, G4, G8, and G12 human rotavirus strains. This finding show that it is possible to carry out the rotavirus infection-protection experiments in mice by using human rotaviruses.