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）（Ｃｒｙｐｔｈｅｃｏｄｉｎｉｕｍｃｏｈｎｉｉ）和特殊涡鞭毛虫尖尾虫（尖尾藻）（Ｏｘｙｒｒｈｉｓｍａｒｉｎａ）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（Ｇｉａｒｄｉａｌａｍｂｌｉａ）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５０ｋＤ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。
when typical dinoflagellate, crypthecodinium cohnii, was detected with immunofluorescent microscopy, its nucleus gave positive reaction to ACA serum against humancentromere / kinetochore proteins. However, in its nucleus, just as in nuclei of Tetrahythena 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 manna, gave the same result which was correspondent with those given by Tetrahymena, Euglena and human Hep-2 cells. With the CENP-B polyclonalantibody, 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 and 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), whileCrypthecodinium 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).The results described above support the proposal that dinoflagellates should be distinguished into two phyla: Phylum oxyrrhinea and Phylum Peridinea (Li, 199o; zhang et al.,1996).
关键词
涡鞭毛虫
着丝粒
动粒蛋白
Major classfication of dinoflagellates, Crypthecodinium cohnii, Oxyrrhismanna, Cetromere/kinetochore proteins, Immuno-blotting
