Ernst Wolfgang, Trummer Evelyn, Mead Jennifer, Bessant Conrad, Strelec Harald, Katinger Hermann, Hesse Friedemann
Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria.
Biotechnol J. 2006 Jun;1(6):639-50. doi: 10.1002/biot.200600010.
Microarray technology for mammalian cells has been utilized mainly for humans, mouse, and rat gene expression analysis. In this approach the feasibility of cross-species hybridization experiments using Chinese hamster ovary (CHO) cells was evaluated. Sequence alignments of available data for CHO were performed against mouse and rat transcripts to determine the homology between the investigated species. We implemented a probability model based on this homology in order to estimate the chance for successful hybridization using Agilent's 60-mer oligonucleotide platform. Heat-shock expression data from CHO, mouse 3T3, and rat A10 cells were generated to determine intraspecies variability, reproducibility, and specificity in order to assess the accuracy of this method. Detected signature genes, in particular from studies with the mouse arrays, showed a reliable similarity between these two rodents and were confirmed by quantitative RT-PCR. Our findings provide evidence that cross-species analysis can be a useful tool to study gene expression profiles of related organisms for which species-specific microarrays are not available.
用于哺乳动物细胞的微阵列技术主要用于人类、小鼠和大鼠的基因表达分析。在这种方法中,评估了使用中国仓鼠卵巢(CHO)细胞进行跨物种杂交实验的可行性。将CHO的可用数据与小鼠和大鼠转录本进行序列比对,以确定所研究物种之间的同源性。我们基于这种同源性实施了一个概率模型,以估计使用安捷伦60-mer寡核苷酸平台成功杂交的机会。生成了来自CHO、小鼠3T3和大鼠A10细胞的热休克表达数据,以确定种内变异性、可重复性和特异性,从而评估该方法的准确性。检测到的特征基因,特别是来自小鼠阵列研究的基因,在这两种啮齿动物之间显示出可靠的相似性,并通过定量RT-PCR得到证实。我们的研究结果提供了证据,表明跨物种分析可以成为研究相关生物体基因表达谱的有用工具,而对于这些生物体,尚无物种特异性微阵列。