Bärlund M, Nupponen N N, Karhu R, Tanner M M, Paavola P, Kallioniemi O P, Kallioniemi A
Laboratory of Cancer Genetics, Institute of Medical Technology, University of Tampere and Tampere University Hospital, Tampere (Finland).
Cytogenet Cell Genet. 1998;82(3-4):189-91. doi: 10.1159/000015096.
Defining boundaries of chromosomal rearrangements at the molecular level would benefit from landmarks that link the cytogenetic map to physical, genetic, and transcript maps, as well as from large-insert FISH probes for such loci to detect numerical and structural rearrangements in metaphase or interphase cells. Here, we determined the locations of 24 genetically mapped CEPH-Mega YACs along the FLpter scale (fractional length from p-telomere) by quantitative fluorescence in situ hybridization analysis. This generated a set of cytogenetically mapped probes for chromosome 17 with an average spacing of about 5 cM. We then developed large-insert YAC, BAC, PAC, or P1 clones to the following 24 known genes, and determined refined map locations along the same FLpter scale: pter-TP53-TOP3-cen-TNFAIP1-ERBB2-TOP2A- BRCA1-TCF11-NME1-HLF-ZNF147/CL N80-BCL5/MPO/SFRS1-TBX2-PECAM1-DDX5/ PRKCA-ICAM2-GH1/PRKAR1A-GRB2-CDK3 /FKHL13-qter. Taken together, these 48 cytogenetically mapped large-insert probes provide tools for the molecular analysis of chromosome 17 rearrangements, such as mapping amplification, deletion, and translocation breakpoints in this chromosome, in cancer and other diseases.
在分子水平上界定染色体重排的边界，将受益于能够把细胞遗传学图谱与物理图谱、遗传图谱及转录图谱联系起来的界标，以及用于此类位点的大插入片段荧光原位杂交（FISH）探针，以检测中期或间期细胞中的数目和结构重排。在此，我们通过定量荧光原位杂交分析确定了24个经遗传定位的CEPH - Mega酵母人工染色体（YAC）沿着FLpter尺度（从p端粒起的分数长度）在17号染色体上的位置。这产生了一组细胞遗传学定位的17号染色体探针，平均间距约为5厘摩（cM）。然后，我们针对以下24个已知基因构建了大插入片段的YAC、细菌人工染色体（BAC）、噬菌体人工染色体（PAC）或P1克隆，并沿着相同的FLpter尺度确定了精确的图谱位置：pter - TP53 - TOP3 - cen - TNFAIP1 - ERBB2 - TOP2A - BRCA1 - TCF11 - NME1 - HLF - ZNF147/CLN80 - BCL5/MPO/SFRS1 - TBX2 - PECAM1 - DDX5/PRKCA - ICAM2 - GH1/PRKAR1A - GRB2 - CDK3/FKHL13 - qter。这些共48个细胞遗传学定位的大插入片段探针共同为17号染色体重排的分子分析提供了工具，比如在癌症及其他疾病中定位该染色体上的扩增、缺失及易位断点。
