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冈比亚按蚊多次吸血后特定mRNA的积累。

The accumulation of specific mRNAs following multiple blood meals in Anopheles gambiae.

作者信息

Nirmala X, Marinotti O, James A A

机构信息

Department of Molecular Biology & Biochemistry, University of California, Irvine, CA 92697-3900, USA.

出版信息

Insect Mol Biol. 2005 Jan;14(1):95-103. doi: 10.1111/j.1365-2583.2005.00535.x.

Abstract

One approach to genetic control of transmission of the parasites that cause human malaria is based on expressing effector genes in mosquitoes that disable the pathogens. Endogenous mosquito promoter and other cis-acting DNA sequences are needed to direct the optimal tissue-, stage- and sex-specific expression of the effector molecules. The mRNA accumulation profiles of eight different genes expressed specifically in the midgut, salivary glands or fat body tissues of the malaria vector, Anopheles gambiae, were characterized as a measure of their suitability to direct the expression of effector molecules designed to disable specific stages of the parasites. RT-PCR techniques were used to determine the abundance of the gene products and their duration following multiple blood meals. Transcription from the midgut-expressed carboxypeptidase-encoding gene, AgCP, follows a cyclical, blood-inducible expression pattern with maximum accumulation every 3 h post blood meal. Other midgut-expressed genes encoding a trypsin and chymotrypsin, Antryp2 and Anchym1, respectively, and the fat body-expressed genes, Vg1 and Cathepsin, also show a blood-inducible pattern of expression with maximum accumulation 24 h after every blood meal. Expression of the Lipophorin gene in the fat body and apyrase and D7-related genes (AgApy and D7r2) in the salivary glands is constitutive and not significantly affected by blood meals. Promoters of the midgut- and fat body-expressed genes may lead to maximum accumulation of antiparasite effector molecule transcripts after multiple blood meals. The multiple feeding behaviour of An. gambiae thus can be an advantage to express high levels of antiparasite effector molecules to counteract the parasites throughout most of adult development.

摘要

一种对导致人类疟疾的寄生虫传播进行基因控制的方法是基于在蚊子中表达能够使病原体失活的效应基因。需要内源性蚊子启动子和其他顺式作用DNA序列来指导效应分子在组织、阶段和性别特异性方面的最佳表达。对在疟疾传播媒介冈比亚按蚊的中肠、唾液腺或脂肪体组织中特异性表达的八个不同基因的mRNA积累谱进行了表征,以此作为衡量它们指导设计用于使寄生虫特定阶段失活的效应分子表达的适用性的指标。使用RT-PCR技术来确定多次进食血液后基因产物的丰度及其持续时间。从中肠表达的羧肽酶编码基因AgCP转录遵循一种周期性的、血液诱导的表达模式,在进食血液后每3小时积累量达到最大值。其他分别编码胰蛋白酶和胰凝乳蛋白酶的中肠表达基因Antryp2和Anchym1,以及脂肪体表达基因Vg1和组织蛋白酶,也显示出血液诱导的表达模式,每次进食血液后24小时积累量达到最大值。脂肪体中脂蛋白基因以及唾液腺中腺苷三磷酸双磷酸酶和D7相关基因(AgApy和D7r2)的表达是组成型的,不受进食血液的显著影响。中肠和脂肪体表达基因的启动子可能会导致多次进食血液后抗寄生虫效应分子转录本的最大积累。冈比亚按蚊的多次进食行为因此可能有利于在成年发育的大部分时间内表达高水平的抗寄生虫效应分子来对抗寄生虫。

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