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Screening of Tissue-Specific Genes and Promoters in Tomato by Comparing Genome Wide Expression Profiles of Arabidopsis Orthologues

机译:通过比较拟南芥直系同源基因全基因组表达谱筛选番茄组织特异性基因和启动子

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摘要

Constitutive overexpression of transgenes occasionally interferes with normal growth and developmental processes in plants. Thus, the development of tissue-specific promoters that drive transgene expression has become agriculturally important. To identify tomato tissue-specific promoters, tissue-specific genes were screened using a series of in silico-based and experimental procedures, including genome-wide orthologue searches of tomato and Arabidopsis databases, isolation of tissue-specific candidates using an Arabidopsis microarray database, and validation of tissue specificity by reverse transcription-polymerase chain reaction (RT-PCR) analysis and promoter assay. Using these procedures, we found 311 tissue-specific candidate genes and validated 10 tissue-specific genes by RT-PCR. Among these identified genes, histochemical analysis of five isolated promoter::GUS transgenic tomato and Arabidopsis plants revealed that their promoters have different but distinct tissue-specific activities in anther, fruit, and root, respectively. Therefore, it appears these in silico-based screening approaches in addition to the identification of new tissue-specific genes and promoters will be helpful for the further development of tailored crop development.
机译:转基因的组成型过表达有时会干扰植物的正常生长和发育过程。因此,驱动转基因表达的组织特异性启动子的开发在农业上变得重要。为了鉴定番茄组织特异性启动子,使用一系列基于计算机模拟和实验的方法筛选了组织特异性基因,包括对番茄和拟南芥数据库进行全基因组直系同源搜索,使用拟南芥微阵列数据库分离组织特异性候选基因,逆转录-聚合酶链反应(RT-PCR)分析和启动子分析验证组织特异性。使用这些程序,我们找到了311个组织特异性候选基因,并通过RT-PCR验证了10个组织特异性基因。在这些鉴定出的基因中,对五种分离的启动子:: GUS转基因番茄和拟南芥植物的组织化学分析表明,它们的启动子分别在花药,果实和根部具有不同但截然不同的组织特异性活性。因此,除了鉴定新的组织特异性基因和启动子外,这些基于计算机模拟的筛选方法似乎将有助于定制作物的进一步发展。

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