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人X染色体DNA的部分纯化与特性分析

Partial purification and characterization of DNA from the human X chromosome.

作者信息

Schmeckpeper B J, Smith K D, Dorman B P, Ruddle F H, Talbot C C

出版信息

Proc Natl Acad Sci U S A. 1979 Dec;76(12):6525-8. doi: 10.1073/pnas.76.12.6525.

Abstract

Human X chromosome DNA was partially purified from a mouse-human hybrid cell line containing a single human chromosome, the X. Enrichment of such DNA was accomplished by two sequential reassociations of radiolabeled hybrid cell DNA with large excesses of mouse DNA. Unreassociated hybrid cell DNA was used as a probe for human X chromosome sequences. The human-specific fraction of probe DNA CONTAINED THREE COMPONENTS. Two of these reassociated to human DNAs at rates proportional to the number of X chromosomes present. These two components were thus localized to the X chromosome. One of these X-specific components, representing about 80% of human-specific probe DNA, consisted of single copy or very low order reiterated DNA. The second X-specific component, representing about 10% of human-specific probe DNA, was about 20-30 times more reiterated. The remaining 10% of human-specific probe DNA, although derived from the X chromosome, reassociated to human DNAs at a rate independent of the number of X chromosomes present. This component was thus homologous to autosomal as well as X chromosome DNA. The probe DNA accounts for approximately half of the human X chromosome, suggesting that the remainder may have homology with mouse DNA.

摘要

从一个含有单条人类染色体X的小鼠 - 人类杂交细胞系中部分纯化了人类X染色体DNA。通过将放射性标记的杂交细胞DNA与大量过量的小鼠DNA进行两次连续复性来实现这种DNA的富集。未复性的杂交细胞DNA用作人类X染色体序列的探针。探针DNA的人类特异性部分包含三个组分。其中两个组分以与存在的X染色体数量成比例的速率与人类DNA复性。因此,这两个组分定位于X染色体。这些X特异性组分中的一个,约占人类特异性探针DNA的80%,由单拷贝或非常低阶的重复DNA组成。第二个X特异性组分,约占人类特异性探针DNA的10%,其重复程度约高20 - 30倍。其余10%的人类特异性探针DNA,虽然源自X染色体,但以与存在的X染色体数量无关的速率与人类DNA复性。因此,该组分与常染色体以及X染色体DNA同源。探针DNA约占人类X染色体的一半,这表明其余部分可能与小鼠DNA具有同源性。

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