Microdissection Based Cloning of a Translocation Breakpoint in a Human Malignant Melanoma
Abstract
Chromosome translocations in human malignancies have identified the genomic location of several important growth-regulatory sequences ( e.g. , cellular oncogenes and suppressor genes). Melanomas are characterized by recurring chromosome alterations, including deletion or translocations of the long arm of chromosome 6 (6q). This report details our efforts to clone the t(l;6)(q21;ql4) breakpoint in a malignant melanoma to further our understanding of the biology of these tumors. The strategy utilized combined microdissection of the translocation chromosome, development and characterization of a DNA microclone library, isolation of cosmids and YACs from the breakpoint region, ordering of clones by two-color metaphase/interphase fluorescence in situ hybridization, and finally, identification of a YAC spanning the translocation breakpoint. By analogy to other tumor systems, molecular examination of the chromosome 6 break point may provide insight into the pathobiology of this important neoplasm 1 To whom requests for reprints should be addressed, at National Center for Human Genome Research/NIH, 49 Convent Drive, MSC 4470, Room 4A22, Bethesda, MD 20892-4470. Phone: (301) 402-2023; Fax: (302) 402-2040.