Grant-in-Aid for international Scientific Research
|Allocation Type||Single-year Grants|
|Research Institution||Yamaguchi University|
FUJISHIMA Masahiro Yamaguchi University, Professor, 理学部, 教授 (40127783)
FOKIN Sergei L St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
SKOBLO Inna I St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
RAUTIAN Maria S St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
OSSIPOV Dmitry V St.-Petersburg State University, Professor, 生物学部, 教授
BRIGGE Theodor Stuttgart University, Assistant, 動物学部, 助手
GORTZ Hans-Dieter Stuttgart University, Professor, 動物学部, 教授
ISHIKAWA Hajime Graduate School of Tokyo University, Professor, 大学院・理学系研究科, 教授 (70012482)
HORI Manabu Yamaguchi University, Assistant, 理学部, 助手 (00253138)
GORT HansーDi シュツッツガルト大学, 生物学部, 教授
|Project Period (FY)
1995 – 1996
Completed(Fiscal Year 1996)
|Budget Amount *help
¥5,800,000 (Direct Cost : ¥5,800,000)
Fiscal Year 1996 : ¥2,700,000 (Direct Cost : ¥2,700,000)
Fiscal Year 1995 : ¥3,100,000 (Direct Cost : ¥3,100,000)
|Keywords||Paramecium / Holospora / Proton pomp / Endosymbiotic bacterium / Endonuclear symbiotic bacterium / Protozoa / Ciliates / Infection / 核内共生細菌 / groEL / ゲノムサイズ / 凍結保存 / モノクローナル抗体|
1.Acidification of inside pH of Paramecium digestive vacuole is induced by a proton pomp (V-ATPase) of the vacuole membrane, and the acidification is indispensble phenomenon for Holospora's escape from the vacuole.
2.Reproductive forms of Holospora obtusa promotes the host cell division not only in low temperature (4ﾟC) but also in 25ﾟC.
3.When the reproductive forms differentiate to the infectious long forms, large amounts of cell-wall antigen (IR-4-1 antigen) are released, or the infectious forms are continuously shedding the antigens, and the shedded antigens stick to the macronuclear envelope and make the envelope unable to pass large molecules such as immunoglobulins inside the macronuclear envelope. This may be a cause for the host death when the macronucleus is filled with the infectious long forms. A mutant strain without the IR-4-1 antigen could not induce the host death even if the host macronucleus was filled with the infectious forms.
4.Genes of stress proteins, groES and groEL homolog, were cloned in H.obtusa.
5.Infectious form-specific 4.5 K protein of H.obtusa was purified and aminoacid sequences was clarified in the total length, and also the gene was cloned. Immunoelectron microscopy with a polyclonal antibody showed that the antigen was localized in the periplasm. Northern blot analysis using the cDNA showed that transcriptio was carried out in intermediate form of the bacterium which appeared in differentiation from the reproductive to the infectious form.
6.Indirect immunofluorescence microscopy and immunoblot with antibodies against Paramecium caudatum actin showed that H.obtusa had a protein with an epitope for the antibody.
Using pulse field gel electrophoresis (PFGE), is was found that genome sizes of H.undulata, H.obtusa and H.recta were 1800,1740 and 1800 kbp respectively.