Crasta Oswald R, Folkerts Otto
CuraGen Corporation, New Haven, CT, USA.
Methods Mol Biol. 2003;236:381-94. doi: 10.1385/1-59259-413-1:381.
The recent rapid developments in genomics tools, technologies, and bioinformatics have revolutionized gene expression analysis. It is now routine to measure gene expression modulation at the genomic level. GeneCalling technology is an open architecture system capable of assaying more than 95% of genes expressed in a tissue. Unlike the closed systems, GeneCalling is not dependent upon an existing sequence or clone database. GeneCalling uses as low as 50 pg of the cDNA from samples and identifies cDNA fragments that are differentially modulated within a set of samples. With the use of 96 pairs of restriction enzymes, more than 30,000 cDNA fragments are routinely assayed to identify those that are differentially modulated. Specific processes, such as SeqCalling, Trace Poisoning, and GeneCall Poisoning, are set up to not only confirm the known genes, but also to clone and analyze unknown and novel genes that have an interesting expression profile. GeneCalling has been successfully applied to expression profiling of several plant and fungal species, and resulted in identification and characterization of genes that are useful in commercial applications towards improving agriculturally important traits in plants.
基因组学工具、技术和生物信息学的最新快速发展彻底改变了基因表达分析。现在，在基因组水平上测量基因表达调控已是常规操作。基因调用技术是一种开放式架构系统，能够检测组织中超过95%的表达基因。与封闭式系统不同，基因调用不依赖于现有的序列或克隆数据库。基因调用使用低至50 pg的样品cDNA，并识别一组样品中差异调控的cDNA片段。通过使用96对限制性酶，通常会检测30000多个cDNA片段，以识别那些差异调控的片段。设置了特定的流程，如序列调用、追踪中毒和基因调用中毒，不仅用于确认已知基因，还用于克隆和分析具有有趣表达谱的未知和新基因。基因调用已成功应用于几种植物和真菌物种的表达谱分析，并导致鉴定和表征了可用于商业应用以改善植物农业重要性状的基因。
