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© 1993 Oxford University Press

OTHER

YAC-assisted cloning of transcribed sequences from the human chromosome 3p21 region

Gina Pengue, Viola Calabrò, Paola Cannada-Bartoli, Pasquale De Luca, Teresa Esposito1, Patricia Taillon-Miller2, Sal LaForgia3, Teresa Druck3, Kay Huebner3, Michele D'Urso1 and Luigi Lania*,

Dipartimento di Genetica, Biologia Generale e Molecolare University of Napoli Via Mezzocannone 8, 80134 Naples 1International Institute of Genetics and Biophysics, CNR Via Marconi 10, 80125 Naples, Italy 2Human Genome Center, Washington University St Louis, MO 63110 3Jefferson Cancer Institute, Thomas Jefferson Medical College Philadelphia, PA 19107, USA

To whom correspondence should be made

Received September 10, 1992; Revised April 8, 1993; Accepted April 8, 1993

The region surrounding the ZNF35 zinc finger protein gene on 3p21 [PDB] is of particular interest, as this region of chromosome 3 is frequently involved in rearrangements and/or deletions associated with various human tumors including lung and renal carcinoma. We have analyzed yeast artificial chromosomes (YACs), identified by PCR screening, using oligonucleotides derived from the ZNF35 gene. PFGE and Southern blot/hybridization analysis revealed that the clones cover 750-kb including the ZNF35 gene. The use of specific somatic cell hybrids have allowed us to locate the YAC contig telomeric to the D3F15S2 locus, in a region which is frequently deleted in lung carcinomas. In addition, we have developed a novel cDNA hybridization protocol allowing the isolation of transcribed sequences present in the overlapping YAC clones. Using the cDNA hybridization selection, we have isolated and characterized one transcribed sequence (D3S1362E) from the 3p21 YAC contig and the corresponding cDNA has been isolated. DNA sequencing analysis indicated that the D3S1363E cDNA codes for a putative transcription factor. Northern blot analysis indicated that the D3S1362E sequence hybridized to multiple transcripts in skeletal muscle, and weakly hybridizing transcripts of similar sizes were detected in other tissues.


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