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Studies on the mechanism of the toxin production in Alexandrium spp.

Studies on the mechanism of the toxin production in Alexandrium spp.
亚历山大藻产毒机制的研究。
批准号:
11660182
负责人:
ISHIMARU Takashi
金额:
$0.58万
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (C)
财政年份:
1999
资助国家:
日本
项目状态:
已结题
起止时间:
1999 至 2000

项目摘要

项目成果

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中文摘要
翻译
为了鉴定麻痹性贝类中毒(麻痹性贝类中毒)产生的相关基因,我尝试利用紫外线和热休克等物理诱变剂,制备麻痹性贝类中毒致病鞭毛藻tamarense Alexandrium、catenella a.m utum a.m utum无毒突变体。紫外辐射处理后用微吸管法分离的塔玛氏A.tamarense亚克隆中,无毒亚克隆的比例很高(78%)。因此,怀疑在最初的克隆中已经存在无毒细胞。然后,从原始克隆培养中分离出51个亚克隆,未经诱变剂处理,证实其中几乎一半的细胞是无毒的,这表明在长时间的批量培养中,至少有一个无毒细胞株是通过自然突变在原始克隆培养中产生的。据推测,除了产毒基因外,无毒亚克隆和有毒亚克隆的基因组之间没有实质性差异。因此,通过比较两个亚克隆的基因组,可以鉴定出该基因。我对两个亚克隆进行了无性系培养,然后按照植物的标准方法尝试提取RNA。然而,由于A.tamarense的RNase活性非常高,所以我没有成功。我优化了RNA分析的样品处理方法,目前正在进行RNA减法杂交分析产生psp的基因。
英文摘要
In order to identify the gene concernig the production of paralytic shellfish poisoning (PSP), I tried to make non-toxic mutants of PSP causative dinoflagellates Alexandrium tamarense, A.catenella and A.minutum by using physical-mutagens such as UV and heat-shock.In the sub-clones of A.tamarense isolated by micoropipet method after the treatment by UV radiation, non-toxic sub-clones appeared in a very high percentage (78%). Therefore, it was suspected that non-toxic cells had already existed in the original clone. Then, 51 sub-clones were isolated without mutagen treatment from the original clone culture, and it was confirmed that almost a half of the cells in that were non-toxic suggesting that at least a single non-toxic cell strain occurred in the original clone culture by a natural mutation in the long duration of batch culture.It is presumed that there is not a substantial difference between the genomes in the non-toxic and toxic sub-clones except in the toxin-producing gene. Thus, by comparing the genomes of the both sub-clones the gene could be identified.I made axenic cultures for the both sub-clones and then tried to extract RNA according to the standard method for plant. However, I could not succeed in it because of very high activity of RNase in A.tamarense. I optimized the sample processing method for the RNA analysis and, currently, the analyses of the PSP-producing gene by using subtractive hybridization of RNA are ongoing.
期刊论文(4)
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科研奖励(0)
会议论文
古谷研,石丸隆,高橋正征: "植物プランクトンの光合成-光曲線の測定"月間海洋. 号外21. 113-122 (2000)
Ken Furuya、Takashi Ishimaru、Masayuki Takahashi:“浮游植物的光合作用 - 光变曲线的测量”海洋月刊 Extra 21. 113-122 (2000)。
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通讯作者:
Koike, K., K.Koike, M.Takagi, T.Ogata and T.Ishimaru: "Evidence of phagotrophy in Dinophysis fortii (Dinophysiales, ), a dinoflagellate that causes diarrhetic shellfish poisoning (DSP)"Phycologycal Research. 48. 121-124 (2000)
Koike, K.、K.Koike、M.Takagi、T.Ogata 和 T.Ishimaru:“Dinophys fortii(Dinophyales,)是一种导致腹泻性贝类中毒 (DSP) 的甲藻中吞噬作用的证据”藻类学研究。
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通讯作者:
Koike,K.,K.Koike,M.Takagi,T.Ogata and T.Ishimaru: "Evidence of phagotrophy in Dinophysis fortii (Dinophysiales, Dinophyceae), a dinoflagellate that causes diarrhetic shellfish poisoning (DSP)"Phycologycal Research. 48. 121-124 (2000)
Koike,K.,K.Koike,M.Takagi,T.Ogata 和 T.Ishimaru:“Dinophys fortii(甲藻目,甲藻科)中吞噬作用的证据,这是一种导致腹泻性贝类中毒 (DSP) 的甲藻”藻类学研究。
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Furuya, K., T.Ishimaru and M.Takahashi: "Measurements of photosynthesis versus light intensity curves in phytoplankton"Gekkan Kaiyo, spec.21, Phytoplankton II. (in Japanese). 116-122 (2000)
Furuya, K.、T.Ishimaru 和 M.Takahashi:“浮游植物光合作用与光强度曲线的测量”Gekkan Kaiyo,spec.21,浮游植物 II。
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A reexamination of the taxonomical study of the genus Skeletonema and their spatial and temporal distribution in Japan
Development of an improving system of bottom hypoxia in estuaries
Studies on the Changes in Environment and Ecosystem in the Antarctic Ocean
Ecosystem and Oceanographic Structure in the Indian Sector of the Antarctic Ocean
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