ISOLATION OF A YEAST ARTIFICIAL CHROMOSOME SPANNING THE 8-21 TRANSLOCATION BREAKPOINT T(8-21)(Q22-Q22.3) IN ACUTE MYELOGENOUS LEUKEMIA
ISOLATION OF A YEAST ARTIFICIAL CHROMOSOME SPANNING THE 8-21 TRANSLOCATION BREAKPOINT T(8-21)(Q22-Q22.3) IN ACUTE MYELOGENOUS LEUKEMIA
复制标题
DOI：
10.1073/pnas.88.11.4882
复制
发表时间：
1991-06-01
影响因子：
11.1
通讯作者：
DRABKIN, HA
中科院分区：
文献类型：
--
作者：
GAO, JZ;ERICKSON, P;DRABKIN, HA
8;易位是急性髓性白血病中最常见的特异性重排之一。通过使用至少三个限制性内切酶（Not I， Sac II和BssHII），我们已经鉴定出位于21号染色体易位断点（21q22.3）两侧的标记（D21S65和NotI边界克隆Not-42，称为探针B）在正常基因组DNA中显示出物理连锁，并且它们之间的距离不超过250-280千碱基。含8的体细胞杂交体DNA的脉冲场凝胶分析21条易位染色体显示了这些标记的重排。一个470千碱基的酵母人工染色体，YAC-Not-42，已被分离，包含这两个探针。利用YAC-Not-42构建的lambda-亚克隆图谱表明，酵母人工染色体DNA的bbb95%（25/26个测试探针）位于断点的近端（D21S65）侧。5例急性髓性白血病患者中期染色体的原位杂交研究21易位证实了这些结果，并证明探针B易位到衍生染色体8。从探针B得到的几乎等于39千碱基的染色体行走，允许鉴定含有衍生染色体8的体细胞杂交DNA的断点。由于探针B含有保守的DNA序列，并且非常接近易位断点，因此它可能代表21号染色体上相关基因的一部分。
The 8;21 translocation is one of the most common specific rearrangements in acute myelogenous leukemia. We have identified markers (D21S65 and a NotI boundary clone, Not-42, referred to as probe B) flanking the chromosome 21 translocation breakpoint (21q22.3) that demonstrate physical linkage in normal genomic DNA, by using at least three restriction endonucleases (Not I, Sac II, and BssHII), and that are located not more than 250-280 kilobases apart. Pulsed-field gel analysis of DNA from somatic cell hybrids containing the 8;21 translocation chromosomes demonstrates rearrangement of these markers. A 470-kilobase yeast artificial chromosome, YAC-Not-42, has been isolated that contains both probes. Mapping of lambda-subclones constructed from YAC-Not-42 suggests that > 95% (25/26 probes tested) of the yeast artificial chromosome DNA is located on the proximal (D21S65) side of the breakpoint. In situ hybridization studies using metaphase chromosomes from five acute myelogenous leukemia patients with the 8;21 translocation confirmed these results and demonstrated the translocation of probe B to the derivative chromosome 8. A chromosome walk of almost-equal-to 39 kilobases from probe B has allowed identification of the breakpoint in DNA from a somatic cell hybrid containing the derivative chromosome 8. Since probe B contains conserved DNA sequences and is in close proximity to the translocation breakpoint, it may represent a portion of the involved gene on chromosome 21.