›› 2010, Vol. 37 ›› Issue (10): 100-104.

• 生物技术 • 上一篇    下一篇

消减cDNA文库差异表达基因检测分析方法的研究进展

岳洋1,王栋1,郝海生1,杜卫华1,赵学明1,朱化彬1,路永强2   

  1. (1中国农业科学院北京畜牧兽医研究所, 北京 100193;2北京市畜牧兽医总站,北京 100107)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-10-20 发布日期:2010-10-20
  • 通讯作者: 王栋

Research Progress on Detection and Assay of Differentially Expressed Genes from Subtractive cDNA Libraries

YUE Yang1,WANG Dong1,HAO Hai-sheng1,DU Wei-hua1,ZHAO Xue-ming1,ZHU Hua-bin1,LU Yong-qiang2   

  1. (1.Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China;2.Beijing General Station of Animal Husbandry and Veterinary Medicine,Beijing 100107,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-10-20 Published:2010-10-20
  • Contact: WANG Dong

摘要: 同已有的差异表达基因分离方法相比,抑制消减杂交以其快速、高效及假阳性率低等优点被广泛应用,并形成了反向Northern斑点杂交和表达谱基因芯片技术2种差减cDNA文库筛选方法,同时,快速发展的生物信息学为文库中大量阳性差异表达片段的序列分析提供了最有力的技术手段,而RT-PCR和实时荧光定量PCR为进一步在mRNA水平验证基因的表达差异性和初步揭示基因功能奠定了技术基础。作者在简要综述上述各方法的原理、特点及应用基础上指出,先测序后筛库的研究策略可以在成本增加较低的前提下大幅度提高差异表达基因的研究效率。

关键词: 文库筛选; 差异表达基因; 检测分析

Abstract: Compared with the existing methods on separating differentially expressed genes, suppression subtractive hybridization has been used extensively for the characteristics of high efficiency and low false positive rate, and reverse Northern dot blotting and expression profile gene chip of two kinds of techniques had been developed to screen the subtractive cDNA libraries. At the same time, the highly developing bioinformatics provides the most forceful means to analyse the sequence information of positive differentially expressed fragments. Furthermore, RT-PCR and real-time fluorescence quantitative PCR are the methods that further validate the differential expression of genes at mRNA level and exposit the gene functions preliminarily. After reviewing the principles, characteristics and applications of these methods, suggestion was given that the efficiency of research on differentially expressed genes will be increased largely while the cost will not if the library screening strategy was used after sequencing the clones.

Key words: library screening; differentially expressed genes; detection and assay

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