Gould R R, Borisy G G
Laboratory of Molecular Biology, University of Wisconsin, Madison 53706.
Exp Cell Res. 1978 May;113(2):369-74. doi: 10.1016/0014-4827(78)90377-4.
The microtubule nucleating capacity of chromosomes was tested in vitro in lysates of Chinese hamster ovary cells. Colcemid-blocked mitotic cells were lysed with the detergent Triton X-100, incubated with exogenous porcine brain tubulin, attached to electron microscope grids and observed as whole-mounts. Under suitable conditions, greater than 98% of the chromosomes gave rise to microtubules at their kinetochore regions, thus unequivocally demonstrating that chromosomes are competent to initiate specifically microtubule formation. The average number of microtubules that polymerized onto a chromosome was 8 +/- 5, and greater than 36% of the chromosomes had between 10 and 19 microtubules per kinetochore region. We conclude that under the lysis conditions employed, virtually all the chromosomes retain their kinetochores, and that the kinetochores retain a substantial fraction of their microtubule nucleating capacity.
在中国仓鼠卵巢细胞裂解物中对染色体的微管成核能力进行了体外测试。用秋水仙酰胺阻断有丝分裂的细胞用去污剂曲拉通X-100裂解，与外源猪脑微管蛋白一起孵育，附着在电子显微镜网格上并作为整装标本观察。在合适的条件下，超过98%的染色体在其动粒区域产生微管，从而明确证明染色体能够特异性地启动微管形成。聚合到一条染色体上的微管平均数量为8±5，超过36%的染色体每个动粒区域有10到19根微管。我们得出结论，在所采用的裂解条件下，几乎所有染色体都保留了它们的动粒，并且动粒保留了相当一部分微管成核能力。
