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Differential expression of Aedes aegypti salivary transcriptome upon blood feeding

机译:埃及伊蚊唾液转录组在采血时的差异表达

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Saliva of Aedes aegypti contains a complex array of proteins essential for both blood feeding and pathogen transmission. A large numbers of those proteins are classified as unknown in regard to their function(s). Understanding the dynamic interactions at the mosquito-host interface can be achieved in part by characterizing mosquito salivary gland gene expression relative to blood feeding. Towards this end, we developed an oligonucleotide microarray representing 463 transcripts to determine differential regulation of salivary gland genes. This microarray was used to investigate the temporal gene expression pattern of Ae. aegypti salivary gland transcriptome at different times post-blood feeding. Expression of the majority of salivary gland genes (77–87%) did not change significantly as a result of blood feeding, while 8 to 20% of genes were down-regulated and 2.8 to 11.6% genes were up-regulated. Up-regulated genes included defensins, mucins and other immune related proteins. Odorant-binding protein was significantly down-regulated. Among unknown function proteins, several were up-regulated during the first three hours post-blood feeding and one was significantly down-regulated. Quantitative real-time RT-PCR was used to substantiate differential expression patterns of five randomly selected genes. Linear regression analysis revealed a high degree of correlation (R2 > 0.89) between oligonucleotide microarray and quantitative RT-PCR data. To our knowledge, this is the first study to investigate differential expression of the Ae. aegypti salivary gland transcriptome upon blood feeding. A microarray provides a robust, sensitive way to investigate differential regulation of mosquito salivary gland genes.
机译:埃及伊蚊的唾液含有一系列复杂的蛋白质,这些蛋白质对于血液供应和病原体传播都是必不可少的。就其功能而言,大量这些蛋白质被分类为未知。了解蚊-宿主界面的动态相互作用可以部分地通过表征蚊唾液腺基因表达相对于采血的特性来实现。为此,我们开发了代表463个转录本的寡核苷酸微阵列,以确定唾液腺基因的差异调控。该微阵列用于研究Ae的瞬时基因表达模式。采食后不同时间的埃及埃及唾液腺转录组。大部分唾液腺基因(77-87%)的表达由于采血而没有显着变化,而8至20%的基因被下调,而2.8至11.6%的基因被上调。上调的基因包括防御素,粘蛋白和其他免疫相关蛋白。气味结合蛋白被显着下调。在未知功能的蛋白质中,有几种在血液喂养后的前三个小时内被上调,而一个被显着下调。实时定量RT-PCR用于证实五个随机选择基因的差异表达模式。线性回归分析显示寡核苷酸微阵列与定量RT-PCR数据之间具有高度相关性(R2> 0.89)。据我们所知,这是第一个研究Ae差异表达的研究。取血时埃及伊蚊唾液腺转录组。微阵列为研究蚊唾液腺基因的差异调控提供了一种鲁棒,灵敏的方法。

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