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Molecular and cellular studies on endonuclear symbionts

核内共生体的分子和细胞研究

基本信息

批准号：
07044202
负责人：
FUJISHIMA Masahiro
金额：
$ 3.71万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

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.

1.质子泵诱导草履虫消化泡内pH酸化2.钝孔孢菌的繁殖型不仅在4 ℃的低温下，而且在25 ℃的低温下都能促进宿主细胞的分裂; 3.当繁殖型分化为侵染型长型时，大量的细胞壁抗原（IR-4-1抗原）被释放，或者感染形式持续地脱落抗原，脱落的抗原粘附在大核包膜上，使包膜不能通过大核包膜内的大分子如免疫球蛋白。这可能是当大核充满传染性长型时宿主死亡的原因。4.克隆了钝头红球藻胁迫蛋白基因groES和groEL同源基因。5.纯化了钝头红球藻侵染型特异性4.5 K蛋白，并对其进行了氨基酸序列分析，克隆了该基因。用多克隆抗体的免疫电镜显示，抗原定位在周质。北方印迹分析表明，波纹草履虫基因组的转录是以中间形式进行的，表现为从生殖型向感染型的分化。6.间接免疫荧光显微镜和抗草履虫肌动蛋白抗体的免疫印迹分析表明，波纹草履虫基因组中存在一个抗草履虫肌动蛋白抗体的表位蛋白。钝头血吸虫和直头血吸虫的核苷酸序列分别为1800、1740和1800 kbp。