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吴传芬, 李靖炎, 代嘉陵, 王永潮. 1996: 涡鞭毛虫(甲藻)着丝粒/动粒蛋白的检查. 动物学研究, 17(3): 307-313.
引用本文: 吴传芬, 李靖炎, 代嘉陵, 王永潮. 1996: 涡鞭毛虫(甲藻)着丝粒/动粒蛋白的检查. 动物学研究, 17(3): 307-313.
WU Chuan-fen, LI Jing-yan, DAI Jia-ling, WANG Yong-chao. 1996. The Detection of Centromere/Kinetochore Proteins in Typical Dinoflagellate and Oxyrrhis. Zoological Research, 17(3): 307-313.
Citation: WU Chuan-fen, LI Jing-yan, DAI Jia-ling, WANG Yong-chao. 1996. The Detection of Centromere/Kinetochore Proteins in Typical Dinoflagellate and Oxyrrhis. Zoological Research, 17(3): 307-313.

涡鞭毛虫(甲藻)着丝粒/动粒蛋白的检查

The Detection of Centromere/Kinetochore Proteins in Typical Dinoflagellate and Oxyrrhis

  • 摘要: 利用ACA血清、抗人着丝粒蛋白B的单抗和多抗、抗CHO细胞动粒蛋白的单抗,对典型涡鞭毛虫隐沟虫(隐甲藻)(Crypthe codinium cohnii)和特殊涡鞭毛虫尖尾虫(尖尾藻)(Oxyrrhis marina)的着丝粒/动粒蛋白进行了检查。用ACA血清作的荧光观察表明,隐沟虫的这些蛋白虽结合在核骨架上,但在间期时并不形成点状的前着丝粒。免疫印迹检查表明两种涡鞭毛虫的着丝粒蛋白B彼此一致,而且与四膜虫和眼虫的也高度一致。但用ACA血清作免疫印迹检查时,尖尾虫的蛋白虽与四膜虫和眼虫的相近,与隐沟虫的却有极大的差异。以抗动粒蛋白的单抗作此种检查时,尖尾虫与眼虫的反应带相同,而隐沟虫则与源真核生物(Archezoa)贾第虫(Giardia lamblia)的相同;而且隐沟虫和贾第虫都与几种原细菌有两条相同的反应带,其中50 kD的一条是尖尾虫和眼虫都没有的。上述发现不仅从一个新的方面支持了认为应把尖尾虫从典型涡鞭毛虫分出来独立为一个门的主张(李靖炎,1990),而且指出典型涡鞭毛虫在后真核生物(Metakaryota)中间是非常原始的。

     

    Abstract: When typical dinoflagellate,Crypthecodinium cohnii,was detected with immuno-fluorescent microscopy,its nucleus gave positive reaction to ACA serum against human centromere/kinetochore proteins.However,in its nucleus,just as in nuclei of Tetrahymena and Euglena,spot-like precentromeres have never been found.The immuno-blotting detections with the monoclonal antibody,mACA-2,against human centromere protein,CENP-B,showed that Crypthecodinium and the special dinoflagellate,Oxyrrhis marina,gave the same result which was correspondent with those given by Tetrahymena,Euglena and human Hep-2 cells.With the CENP-B polyclonal antibody,ra-ACA-2,the two dinoflagellates,other protists and Hep-2 cells also gave the results identical to each.These results indicated that CENP-B was highly conservative in eukaryotic evolution.In fact,all the centromere/kinetochore proteins are quite conservative.Therefore,in the immuno-blotting detections done with ACA serum,the positive bands of Tetrahymena and Euglena are quite similar to those of human Hep-2 cells.The bands of the special dinoflagellate,Oxyrrhis,are similar to other protists.although a few bands are absent.However,the bands of typical dinoflagellate are obviously different from Oxyrrhis Sand other eukaryotes.It means that two dinoflagellates are far from each other.The results obtained with the monoclonal antibody,mAb37A5,against the kinetochore proteins of CHO cells,also proposed the above suggestion.In the detections,Tetrahymena gave only a 120 kD band; Euglena and Oxyrrhis gave two bands (45 kD and 120 kD),while Crypthecodinium and archezoan,Giardia,gave three bands (45 kD,50 kD and 120 kD).It is very interesting that various archaebacteria all gave the 45 kD band and the 50 kD band (the third band is of 40 kD)(Wu et al.,1996b).

     

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