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对灵长类有丝分裂染色体具有特异性的单克隆抗体。

Monoclonal antibody with specificity to mitotic chromosomes of primates.

作者信息

Davis F M, Wegner R D, Rao P N

出版信息

Exp Cell Res. 1987 Jun;170(2):417-27. doi: 10.1016/0014-4827(87)90317-x.

Abstract

Fusion of a cell in mitosis with a cell in interphase results in the condensation of chromatin in the interphase nucleus into chromosomes. Premature chromosome condensation is caused by certain proteins, called mitotic factors, that are present in the mitotic cell and are localized on chromosomes. Extracts from mitotic cells were used to immunize mice to produce monoclonal antibodies specific for cells in mitosis. Among the antibodies obtained, the MPM-4 antibody defines a 125-kD polypeptide antigen located on mitotic chromosomes by indirect immunofluorescence. Although the polypeptide antigen is present in approximately equal concentrations in extracts of interphase cells and mitotic cells, as revealed by immunoblots, it cannot be detected cytologically in the former. Cell fractionation experiments showed that the 125-kD antigen is found in the cytoplasm of interphase cells and metaphase cells, but is concentrated in fractions containing metaphase chromosomes, although not detectable in interphase nuclei. Even though the antigen is apparently primate-specific, it binds to mitotic chromosomes and prematurely condensed chromosomes in human-rodent cell hybrids without regard to the species of origin of the mitotic inducer. The presence of the antigen in the cytoplasm of interphase cells and the chromosomes of mitotic cells suggests a relationship between the presence of the antigen on chromosomes and the process of chromosome condensation and decondensation.

摘要

处于有丝分裂期的细胞与处于间期的细胞融合,会导致间期细胞核中的染色质凝聚成染色体。早熟染色体凝聚是由某些称为有丝分裂因子的蛋白质引起的,这些蛋白质存在于有丝分裂细胞中并定位于染色体上。用有丝分裂细胞提取物免疫小鼠,以产生对有丝分裂期细胞特异的单克隆抗体。在所获得的抗体中,MPM - 4抗体通过间接免疫荧光确定了一种位于有丝分裂染色体上的125kD多肽抗原。虽然免疫印迹显示该多肽抗原在间期细胞提取物和有丝分裂细胞提取物中的浓度大致相等,但在间期细胞中无法通过细胞学方法检测到。细胞分级分离实验表明,125kD抗原存在于间期细胞和中期细胞的细胞质中,但集中在含有中期染色体的级分中,尽管在间期核中无法检测到。即使该抗原显然是灵长类特异性的,但它能与人类 - 啮齿动物细胞杂种中的有丝分裂染色体和早熟凝聚染色体结合,而不考虑有丝分裂诱导剂的来源物种。该抗原在间期细胞的细胞质和有丝分裂细胞的染色体中的存在表明,染色体上抗原的存在与染色体凝聚和解聚过程之间存在关联。

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