Zhang J, Trent J M, Meltzer P S
Department of Radiation Oncology, University of Michigan, Ann Arbor 48109-0668.
Oncogene. 1993 Oct;8(10):2827-31.
We describe a novel strategy characterizing gene amplification in human neoplasia based on targeting double minutes (dmin) and homogeneously staining regions (hsr) for chromosome microdissection. This strategy allows the rapid generation of an amplification unit microclone library and permits the rapid identification of the chromosomal origin of the amplified sequences following fluorescence in situ hybridization (FISH). This strategy has been applied to an hsr-bearing malignant melanoma cell line, HA-A, which was then demonstrated to encode multiple overexpressed copies of the IGF1R gene. This strategy combines all steps for detection, cloning, mapping and isolation of amplified gene(s) into a single process that is readily applicable to any specimen carrying cytologic evidence of gene amplification.
我们描述了一种基于靶向双微体（dmin）和均匀染色区（hsr）进行染色体显微切割来鉴定人类肿瘤中基因扩增的新策略。该策略能够快速构建扩增单元微克隆文库，并在荧光原位杂交（FISH）后快速鉴定扩增序列的染色体起源。此策略已应用于携带hsr的恶性黑色素瘤细胞系HA-A，结果表明该细胞系编码多个IGF1R基因的过表达拷贝。该策略将扩增基因的检测、克隆、定位和分离的所有步骤整合到一个单一过程中，该过程可轻松应用于任何携带基因扩增细胞学证据的标本。
