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机构地区:[1]浙江大学原子核农业科学研究所,杭州310029
出 处:《中国生物化学与分子生物学报》2002年第3期259-264,共6页Chinese Journal of Biochemistry and Molecular Biology
摘 要:自 90年代早期发展以来 ,差异基因表达 (DGE)技术在许多领域得到了应用 .“开放”结构系统的DGE技术不需原始的生物学或序列信息 ,而且可应用于任何种群 .主要介绍 6项开放的DGE技术 :cDNA代表性差示分析 (cDNA RDA)、基因表达系统分析 (SAGE)、表达序列标签串联排列连接(TALEST) ,和早期的DGE技术差异显示 (DD)、随机引物聚合酶链反应 (AP PCR) ,以及一项受专利保护的技术———GeneCalling .通过几项重要的参数对这些技术进行了比较 ,认为DD虽然有其致命的弱点 ,但在目前仍然应用得非常广泛 .cDNA RDA能有效富增特异片段 ,扣除共有序列 ,如能和SAGE结合 ,将能进一步促进其发展 .TALEST和GeneCalling操作较简便 ,一次试验能获得大量的数据 ,但是分析这些数据比较麻烦 ,须借助另外的分析软件 .最后介绍了应用DGE技术取得的最新成果 .Differential gene expression (DGE) technologies have been applied in various fields since their development in the early 1990s. `Open' DGE technologies, which do not need pre biological or sequence information,could be applied to any organisms. Six ‘open' DGE technologies are introduced: cDNA representational difference analysis(cDNA RDA),serial analysis of gene expression (SAGE),tandem arrayed ligation of expressed sequence tags(TALEST),differential display(DD),arbitrarily primed polymerase chain reaction(AP PCR) and GeneCalling. At present, DD is applied widely in despite of its high rate of false positives. cDNA RDA is one of the earliest well established technologies for DGE analysis. A drawback of cDNA RDA is technically strict demanding. An adaptation of cDNA RDA with features incorporated from SAGE is termed integrated procedure for gene identification (IPGI). TALEST was recently described and is unique among expression technologies in that it does not rely on PCR amplification to generate the initial tag arrays. Analysis of TALEST data is facilitated through web browser based software. In GeneCalling cDNA fragments derived from differentially expressed genes are automatically identified by electronically comparing relative intensity of each peak. Simultaneously the fragment information is queried against a species specific database using the precise size of the fragment and the fragment flanking sequence. The newest applications of these technologies are described at last.
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