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首页> 外文期刊>Genomics >SAGE tag based cDNA microarray analysis during larval to pupal development and isolation of novel cDNAs in Bombyx mori.
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SAGE tag based cDNA microarray analysis during larval to pupal development and isolation of novel cDNAs in Bombyx mori.

机译：在家蚕幼虫发育和分离过程中，基于SAGE标签的cDNA微阵列分析。

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Many genes act together during the complex process of insect larval and pupal development. 20-hydroxyecdysone interacts with juvenile hormone to control insect growth and development and then activates several transcription factors, i.e., Broad, E74, and E75, and, subsequently, the late target genes. To investigate this phenomenon, we used serial analysis of gene expression (SAGE) tag-based cDNA microarray analysis to monitor the global gene expression profile during larval development and larva-pupa metamorphosis of the silkworm Bombyx mori. Of the 330 clones that were dotted to the chip, 267 were obtained by generating longer cDNA fragments from SAGE tags for gene identification, and the others were obtained from SAGE tag-matched genes or expressed sequence tags from public databases. According to the gene expression profile, the genes were classified into 12 clusters using a self-organizing map analysis. The results were partially confirmed using real-time reverse transcription-polymerase chain reaction. We obtained 22 full-length cDNAs using rapid amplification of 5' cDNA ends, of which eight genes were novel in the silkworm. Our results indicated that use of a cDNA microarray based on SAGE tags is effective for identifying and examining some low-expression genes associated with insect development.
机译：在昆虫幼虫和and发育的复杂过程中，许多基因共同起作用。 20-羟基蜕皮激素与少年激素相互作用以控制昆虫的生长和发育，然后激活几个转录因子，即Broad，E74和E75，以及随后的晚期靶基因。为了研究此现象，我们使用了基于基因表达（SAGE）标签的序列分析基于cDNA的微阵列分析，来监测家蚕Bombyx mori的幼虫发育和幼虫pu变态期间的全局基因表达谱。在点缀到芯片上的330个克隆中，有267个是通过从SAGE标签生成更长的cDNA片段以进行基因鉴定而获得的，其余的则是从SAGE标签匹配的基因或从公共数据库中表达的序列标签中获得的。根据基因表达谱，使用自组织图谱分析将基因分为12个簇。使用实时逆转录-聚合酶链反应部分证实了该结果。我们使用5'cDNA末端的快速扩增获得了22个全长cDNA，其中8个基因在蚕中是新的。我们的结果表明，使用基于SAGE标签的cDNA微阵列可有效地识别和检查与昆虫发育相关的一些低表达基因。

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