Gimbel W, Klein V, Brass N, Fischer U, Piontek K, Overmyer K, Göttert E, Zang K D, Meese E
Department of Human Genetics, Medical School, University of Saarland, Homburg/Saar, Germany.
Cytogenet Cell Genet. 1995;71(1):81-5. doi: 10.1159/000134068.
Recently, we reported the generation of a heteronuclear (hn) cDNA library from a human x rodent somatic cell hybrid retaining human chromosome 22. Here, we report the characterization and localization of 12 cDNA derived clones from this library. Human-specific cDNA sequences have been selectively amplified by inter-Alu PCR. To exclude Alu transcripts, only clones with inserts larger than 500 bp were analyzed. Ten of the 12 clones were localized by PCR on chromosome 22, with four clones mapping on additional chromosomes. One PCR-localized clone and two additional clones were mapped on chromosome 22 by in situ hybridization. Transcripts in human cells were identified for seven of the eight clones analyzed by RT-PCR. None of the clones showed significant sequence similarities within the GenBank and EMBL databases, indicating that these clones represent previously unknown genes. This is the first report on the isolation of chromosome 22-specific transcripts from a human x rodent somatic cell hybrid.
最近,我们报道了从保留人类22号染色体的人-啮齿动物体细胞杂种中构建异核(hn)cDNA文库。在此,我们报道了来自该文库的12个cDNA衍生克隆的特征和定位。通过Alu间PCR选择性扩增了人类特异性cDNA序列。为排除Alu转录本,仅分析插入片段大于500 bp的克隆。12个克隆中的10个通过PCR定位在22号染色体上,4个克隆定位在其他染色体上。通过原位杂交将1个PCR定位的克隆和另外2个克隆定位在22号染色体上。通过RT-PCR分析的8个克隆中有7个在人类细胞中鉴定出转录本。在GenBank和EMBL数据库中,这些克隆均未显示出明显的序列相似性,表明这些克隆代表了以前未知的基因。这是关于从人-啮齿动物体细胞杂种中分离22号染色体特异性转录本的首次报道。