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Differential gene expression in abdomens of the malaria vector mosquito, Anopheles gambiae, after sugar feeding, blood feeding and Plasmodium berghei infection

机译:喂糖,喂血和伯氏疟原虫感染后,疟疾媒介蚊子冈比亚按蚊腹部的差异基因表达

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Background Large scale sequencing of cDNA libraries can provide profiles of genes expressed in an organism under defined biological and environmental circumstances. We have analyzed sequences of 4541 Expressed Sequence Tags (ESTs) from 3 different cDNA libraries created from abdomens from Plasmodium infection-susceptible adult female Anopheles gambiae. These libraries were made from sugar fed (S), rat blood fed (RB), and P. berghei-infected (IRB) mosquitoes at 30 hours after the blood meal, when most parasites would be transforming ookinetes or very early oocysts. Results The S, RB and IRB libraries contained 1727, 1145 and 1669 high quality ESTs, respectively, averaging 455 nucleotides (nt) in length. They assembled into 1975 consensus sequences – 567 contigs and 1408 singletons. Functional annotation was performed to annotate probable molecular functions of the gene products and the biological processes in which they function. Genes represented at high frequency in one or more of the libraries were subjected to digital Northern analysis and results on expression of 5 verified by qRT-PCR. Conclusion 13% of the 1965 ESTs showing identity to the A. gambiae genome sequence represent novel genes. These, together with untranslated regions (UTR) present on many of the ESTs, will inform further genome annotation. We have identified 23 genes encoding products likely to be involved in regulating the cellular oxidative environment and 25 insect immunity genes. We also identified 25 genes as being up or down regulated following blood feeding and/or feeding with P. berghei infected blood relative to their expression levels in sugar fed females.
机译:背景技术cDNA文库的大规模测序可以提供在确定的生物学和环境条件下在生物体中表达的基因的概况。我们已经分析了从疟原虫感染易感成年雌性冈比亚按蚊的腹部创建的3个不同cDNA文库中的4541个表达序列标签(EST)的序列。这些文库是在食血后30小时由食糖(S),食鼠血(RB)和伯氏疟原虫感染(IRB)蚊子制成的,那时大多数寄生虫会转化成卵形或很早的卵囊。结果S,RB和IRB文库分别包含1727、1145和1669个高质量EST,平均长度为455个核苷酸(nt)。他们组装成1975个共有序列-567个重叠群和1408个单例。进行功能注释以注释基因产物的可能分子功能及其在其中起作用的生物学过程。对一个或多个文库中以高频率表示的基因进行数字Northern分析,并通过qRT-PCR验证5的表达结果。结论1965年的EST中有13%表现出与冈比亚曲霉基因组序列相同的特性,这些基因代表新基因。这些以及许多EST上存在的非翻译区(UTR),将为进一步的基因组注释提供信息。我们已经确定了23种编码可能参与调节细胞氧化环境的产物的基因,以及25种昆虫免疫基因。我们还确定了25个基因在喂食和/或喂食伯氏疟原虫感染的血液后相对于其在食糖的雌性动物中的表达水平上调或下调。

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