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首页> 外文学位 >Development of expressed sequence tags (ESTS) and cDNA microarrays for analysis of differentially expressed genes in the channel catfish (Ictalurus punctatus) brain.
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Development of expressed sequence tags (ESTS) and cDNA microarrays for analysis of differentially expressed genes in the channel catfish (Ictalurus punctatus) brain.

机译：开发表达序列标签（ESTS）和cDNA微阵列，以分析海the鱼（Ictalurus punctatus）脑中差异表达的基因。

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摘要

The objectives of this research were to develop expressed sequence tags from the channel catfish (Ictalurus punctatus) brain and to develop microarray technologies for analysis of differentially expressed genes in the brain in response to cold acclimation. Expressed sequence tag (EST) analysis was conducted using a cDNA library made from the brain mRNA of channel catfish. ESTs were used to develop microarray technology to determine differentially expressed genes to reveal molecular mechanisms of cold acclimation. A total 1,201 brain cDNA clones that represented 660 unique genes were analyzed for transcriptome analysis in catfish to develop molecular reagents for cDNA microarray analysis to reveal molecular mechanisms of cold acclimation. Of the 660 unique genes, 330 clones were identified as known genes by BLAST searches and 330 clones as unknown genes.; Among 660 analyzed genes, 62 were differentially expressed at 12°C as compared to 24°C. The gene induction upon temperature shift was rapid within two hours. The major categories of differentially expressed genes include: (1) chaperones such as Hsp70 and Hsp70/Hsp90 organizing protein, (2) transcription factors and genes involved in signal transduction pathways such as zinc finger proteins, calmodulin kinase inhibitor, the nuclear autoantigen SG2NA, interferon regulatory factor 3, and inorganic pyrophosphatase, (3) genes involved in lipid metabolism such as TB2 and acyl CoA binding protein, and (4) genes involved in the translational machinery such as ribosomal proteins. Some genes were induced transiently, whereas others were induced in a durable fashion. Several genes were down regulated among which were mainly ribosomal protein genes, indicating reduced metabolic activity with extended incubation at the low temperature. This research indicated that catfish respond to low temperature by adjusting expression of a large number of genes. The rapid induction of proteins involved in signal transductions and chaperones suggest that both de novo synthesis of cold-induced proteins and modification of existing proteins are required in adaptation and tolerance of fish to low temperature environment.
机译：这项研究的目的是从from鱼的大脑中开发表达的序列标签，并开发微阵列技术来分析冷适应后大脑中差异表达的基因。表达的序列标签（EST）分析是使用由channel鱼的脑mRNA制成的cDNA文库进行的。 EST用于开发微阵列技术，以确定差异表达的基因以揭示冷驯化的分子机制。分析了代表660个独特基因的总共1,201个脑cDNA克隆，用于cat鱼的转录组分析，以开发用于cDNA微阵列分析的分子试剂，揭示了冷驯化的分子机制。在660个独特基因中，通过BLAST搜索将330个克隆鉴定为已知基因，将330个克隆鉴定为未知基因。在660个分析的基因中，与24°C相比，在12°C有62个差异表达。温度改变后的基因诱导在两个小时内迅速。差异表达基因的主要类别包括：（1）伴侣蛋白（例如Hsp70和Hsp70 / Hsp90组织蛋白），（2）转录因子和参与信号转导途径的基因，例如锌指蛋白，钙调蛋白激酶抑制剂，核自身抗原SG2NA，干扰素调节因子3和无机焦磷酸酶，（3）参与脂质代谢的基因，例如TB2和酰基CoA结合蛋白，以及（4）参与翻译机制的基因，例如核糖体蛋白。一些基因被瞬时诱导，而另一些则以持久的方式被诱导。几个基因被下调，其中主要是核糖体蛋白基因，表明在低温下扩展孵育会降低代谢活性。这项研究表明cat鱼通过调节大量基因的表达来应对低温。快速诱导参与信号转导和伴侣蛋白的蛋白质表明，冷诱导蛋白质的 de novo 合成和现有蛋白质的修饰都是鱼类对低温环境的适应性和耐受性所必需的。

著录项

  • 作者

    Ju, Zhenlin.;

  • 作者单位

    Auburn University.;

  • 授予单位 Auburn University.;
  • 学科 Biology Molecular.; Agriculture Fisheries and Aquaculture.
  • 学位 Ph.D.
  • 年度 2001
  • 页码 79 p.
  • 总页数 79
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;水产、渔业;
  • 关键词

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