YAC-ASSISTED CLONING OF TRANSCRIBED SEQUENCES FROM THE HUMAN CHROMOSOME-3P21 REGION

The region surrounding the ZNF35 zinc finger protein gene on 3p21 is o f particular interest, as this region of chromosome 3 is frequently in volved in rearrangements and/or deletions associated with various huma n tumors including lung and renal carcinoma. We have analyzed yeast ar tificial chromosomes (YACs), identified by PCR screening, using oligon ucleotides derived from the ZNF35 gene. PFGE and Southern blot/hybridi zation analysis revealed that the clones cover 750-kb including the ZN F35 gene. The use of specific somatic cell hybrids have allowed us to locate the YAC contig telomeric to the D3F15S2 locus, in a region whic h is frequently deleted in lung carcinomas. In addition, we have devel oped a novel cDNA hybridization protocol allowing the isolation of tra nscribed sequences present in the overlapping YAC clones. Using the cD NA hybridization selection, we have isolated and characterized one tra nscribed sequence (D3S1362E) from the 3p21 YAC contig and the correspo nding cDNA has been isolated. DNA sequencing analysis indicated that t he D3S1363E cDNA codes for a putative transcription factor. Northern b lot analysis indicated that the D3S1362E sequence hybridized to multip le transcripts in skeletal muscle, and weakly hybridizing transcripts of similar sizes were detected in other tissues.