Dowdy S F, Scanlon D J, Fasching C L, Casey G, Stanbridge E J
Department of Microbiology and Molecular Genetics, University of California, Irvine 92727.
Genes Chromosomes Cancer. 1990 Nov;2(4):318-27. doi: 10.1002/gcc.2870020410.
The microcell-mediated chromosome transfer technique has been used to introduce whole chromosomes into malignant cells and revert the tumorigenic phenotype. However, in most instances the limited availability of selectable chromosomes has hindered the ability to reduce the region containing the tumor suppressive information. The work presented here describes a new method to enrich for specific chromosomal arm deletions of selectable chromosomes and thereby more finely focus upon the genetic region of interest. The irradiation-microcell mediated chromosome transfer (XMMCT) technique involves the irradiation of microcells containing single human chromosomes followed by fusion to a nonirradiated host and cytogenetic characterization. The XMMCT procedure was performed on a microcell hybrid containing a der(11) as the only human chromosome. The resultant irradiated microcell hybrids were found to have deletions that ranged from simple interstitial deletions to complex deletions/rearrangements involving only the human der(11) chromosome. The XMMCT procedure has broad applications in generating chromosomal reagents for mapping genetic loci and for use in functional analyses such as tumor suppression studies.
微细胞介导的染色体转移技术已被用于将整条染色体导入恶性细胞并逆转致瘤表型。然而,在大多数情况下,可选择染色体的可用性有限,阻碍了缩小包含肿瘤抑制信息区域的能力。本文介绍的工作描述了一种富集可选择染色体特定染色体臂缺失的新方法,从而更精确地聚焦于感兴趣的遗传区域。辐射-微细胞介导的染色体转移(XMMCT)技术包括对含有单条人类染色体的微细胞进行辐射,然后与未辐射的宿主细胞融合并进行细胞遗传学表征。XMMCT程序是在一个含有der(11)作为唯一人类染色体的微细胞杂种上进行的。结果发现,产生的辐射微细胞杂种具有从简单的中间缺失到仅涉及人类der(11)染色体的复杂缺失/重排等各种缺失。XMMCT程序在生成用于绘制遗传位点的染色体试剂以及用于诸如肿瘤抑制研究等功能分析方面具有广泛的应用。