Isolation And Cloning Of Expressed Sequences Within A Region Flanking ANK1 On Chromosome 8p11.2

Cancer is one of major causes of death in men. Despite its high incidence, little is known regarding the molecular pathogenesis of most cancers. It is thought that multiple genetic changes are associated with the initiation and progression of cancer, such as the activation of oncogenes and inactivation of tumor suppressor genes (TSGs). LOH (loss of heterozygosity) analysis, in which deletion of one allele is detected in tumor cells of a heterozygous individual, has been used to search for the loss of TSGs throughout the human genome. LOH at the ANK1 locus or in the ANK1-D8S255 region of chromosome band 8p11.2 has frequently been observed in several types of human cancer, including prostatic, colorectal, head and neck carcinomas. It has been postulated that loss or inactivation of a TSG near the ANK1 locus or in the ANK1-D8S255 region may play an important role in progression of these tumors. In addition, more than ten cases of an atypical myeloproliferative disorder were reported, in which the leukemia cells have the translocation involving 8p11. Other disease genes which may be located on 8p11 or 8p such as an oncogene related to breast cancer and a gene responsible for congenital heart diseases have not been cloned so far. Our studies mainly included the isolation of single-copy sequences from the region flanking ANK1 locus, screening of CEPH yeast artificial chromosome clones (YACs) with the single-copy sequence probe identified, isolation of expressed sequences from the identified YACs by cDNA selection, and screening of the arrayed fetal brain cDNA library with the probe selected from the isolated expressed sequences. Our efforts were to provide the basis for the positional cloning of these genes or the identification of the mutant genes using the positional candidate strategy.