Pengue G, Calabrò V, Cannada-Bartoli P, De Luca P, Esposito T, Taillon-Miller P, LaForgia S, Druck T, Huebner K, D'Urso M
Dipartimento di Genetica, Biología Generale e Molecolare, University of Napoli, Italy.
Hum Mol Genet. 1993 Jun;2(6):791-6. doi: 10.1093/hmg/2.6.791.
The region surrounding the ZNF35 zinc finger protein gene on 3p21 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.
位于3p21的ZNF35锌指蛋白基因周围区域尤其令人关注，因为3号染色体的这个区域经常参与与包括肺癌和肾癌在内的各种人类肿瘤相关的重排和/或缺失。我们分析了通过PCR筛选鉴定的酵母人工染色体（YAC），使用的寡核苷酸源自ZNF35基因。脉冲场凝胶电泳（PFGE）和Southern印迹/杂交分析表明，这些克隆覆盖了包括ZNF35基因在内的750kb区域。使用特定的体细胞杂种使我们能够将YAC重叠群定位到D3F15S2基因座的端粒，该区域在肺癌中经常缺失。此外，我们开发了一种新颖的cDNA杂交方案，可分离重叠YAC克隆中存在的转录序列。通过cDNA杂交筛选，我们从3p21 YAC重叠群中分离并鉴定了一个转录序列（D3S1362E），并分离出了相应的cDNA。DNA测序分析表明，D3S1363E cDNA编码一种假定的转录因子。Northern印迹分析表明，D3S1362E序列与骨骼肌中的多个转录本杂交，并且在其他组织中检测到大小相似的弱杂交转录本。
