High-throughput GLGI procedure for converting a large number of serial analysis of gene expression tag sequences into 3′ complementary DNAs
High-throughput GLGI procedure for converting a large number of serial analysis of gene expression tag sequences into 3′ complementary DNAs
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DOI：
10.1002/gcc.10017
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发表时间：
2002-03-01
影响因子：
3.7
通讯作者：
Wang, SM
中科院分区：
文献类型：
--
作者：
Chen, JJ;Lee, SG;Wang, SM
基因表达系列分析（SAGE）是一种全基因组基因表达分析的有力技术。然而，由于两个原因，三分之二的SAGE标签不能直接用于基因鉴定。首先，由于SAGE标签序列的长度短，许多SAGE标签匹配几个已知的表达序列。其次，许多SAGE标签与任何已知的表达序列不匹配，可能是因为与这些SAGE标签对应的序列尚未被鉴定。这两个问题可以通过使用GLGI技术（从SAGE标签产生更长的cDNA片段用于基因鉴定）将SAGE标签延伸到3 ′互补DNA（cDNA）中来解决。我们改进了原来的GLGI技术，使之成为一种高通量的方法，可以同时将大量的SAGE标签转化为相应的3'cDNA。整个过程是简单，快速，低成本，高效率，我们使用这个程序分析了数百个SAGE标签。除了鉴定具有多个匹配的SAGE标签的正确基因之外，GLGI还可以通过将新的SAGE标签转化为3'cDNA来用于大规模鉴定新基因。应用这种高通量的程序应加快在人类和其他真核生物基因组中的基因识别的速度显着。(C)2002 Wiley-Liss，Inc.
Serial analysis of gene expression (SAGE) is a powerful technique for genome-wide analysis of gene expression. However, two-thirds of SAGE tags cannot be used directly for gene identification for two reasons. First, many SAGE tags match several known expressed sequences, owing to the short length of SAGE tag sequences. Second, many SAGE tags do not match any known expressed sequences, presumably because the sequences corresponding to these SAGE tags have not been identified. These two problems can be solved by extension of the SAGE tags into 3' complementary DNAs (cDNAs) by use of the GLGI technique (generation of longer cDNA fragments from SAGE tags for gene identification). We have improved the original GLGI technique into a high-throughput procedure for simultaneous conversion of a large number of SAGE tags into corresponding 3' cDNAs. The whole process is simple, rapid, low-cost, and highly efficient, as shown by our use of this procedure for analyzing hundreds of SAGE tags. In addition to identifying the correct gene for SAGE tags with multiple matches, GLGI can be used for large-scale identification of novel genes by converting novel SAGE tags into 3' cDNAs. Applying this high-throughput procedure should accelerate the rate of gene identification significantly in the human and other eukaryotic genomes. (C) 2002 Wiley-Liss, Inc.