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Gene Organization in Rice Revealed by Full-Length cDNA Mapping and Gene Expression Analysis through Microarray

机译:全长cDNA图谱显示水稻基因组织和微阵列基因表达分析

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摘要

Rice (Oryza sativa L.) is a model organism for the functional genomics of monocotyledonous plants since the genome size is considerably smaller than those of other monocotyledonous plants. Although highly accurate genome sequences of indica and japonica rice are available, additional resources such as full-length complementary DNA (FL-cDNA) sequences are also indispensable for comprehensive analyses of gene structure and function. We cross-referenced 28.5K individual loci in the rice genome defined by mapping of 578K FL-cDNA clones with the 56K loci predicted in the TIGR genome assembly. Based on the annotation status and the presence of corresponding cDNA clones, genes were classified into 23K annotated expressed (AE) genes, 33K annotated non-expressed (ANE) genes, and 5.5K non-annotated expressed (NAE) genes. We developed a 60mer oligo-array for analysis of gene expression from each locus. Analysis of gene structures and expression levels revealed that the general features of gene structure and expression of NAE and ANE genes were considerably different from those of AE genes. The results also suggested that the cloning efficiency of rice FL-cDNA is associated with the transcription activity of the corresponding genetic locus, although other factors may also have an effect. Comparison of the coverage of FL-cDNA among gene families suggested that FL-cDNA from genes encoding rice- or eukaryote-specific domains, and those involved in regulatory functions were difficult to produce in bacterial cells. Collectively, these results indicate that rice genes can be divided into distinct groups based on transcription activity and gene structure, and that the coverage bias of FL-cDNA clones exists due to the incompatibility of certain eukaryotic genes in bacteria.
机译:水稻(Oryza sativa L.)是单子叶植物功能基因组学的典范生物,因为其基因组大小明显小于其他单子叶植物。尽管可获得highly稻和粳稻的高度精确的基因组序列,但是对于全面分析基因结构和功能,诸如全长互补DNA(FL-cDNA)序列之类的其他资源也是必不可少的。我们交叉引用了通过映射578K FL-cDNA克隆与TIGR基因组大会中预测的56K基因座而定义的水稻基因组中的28.5K基因座。根据注释状态和相应cDNA克隆的存在,将基因分为23K注释的表达(AE)基因,33K注释的非表达(ANE)基因和5.5K注释的非表达(NAE)基因。我们开发了一种60mer寡核苷酸阵列,用于分析每个基因座的基因表达。基因结构和表达水平的分析表明,NAE和ANE基因的基因结构和表达的一般特征与AE基因有很大不同。结果还表明,水稻FL-cDNA的克隆效率与相应遗传基因座的转录活性有关,尽管其他因素也可能有影响。基因家族中FL-cDNA覆盖率的比较表明,在细菌细胞中很难产生来自编码水稻或真核生物特异结构域的基因的FL-cDNA,以及涉及调节功能的基因。总的来说,这些结果表明水稻基因可以根据转录活性和基因结构分为不同的组,并且由于某些真核基因在细菌中的不相容性,存在着FL-cDNA克隆的覆盖偏差。

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