Identification of genes involved in interactions between Biomphalaria glabrata and Schistosoma mansoni by suppression subtractive hybridization
Identification of genes involved in interactions between Biomphalaria glabrata and Schistosoma mansoni by suppression subtractive hybridization
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DOI：
10.1016/j.molbiopara.2006.09.009
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发表时间：
2007-01-01
影响因子：
1.5
通讯作者：
Jones, Catherine S.
中科院分区：
文献类型：
--
作者：
Lockyer, Anne E.;Spinks, Jennifer;Jones, Catherine S.
光滑双脐螺是曼氏血吸虫（一种医学上重要的血吸虫）的中间蜗牛宿主。为了鉴定参与蜗牛-血吸虫相互作用的转录本，使用抑制消减杂交（SSH）在暴露于寄生虫的抗血吸虫菌株和光滑拟杆菌之间以及暴露于血吸虫和未暴露的来自抗性蜗牛系的蜗牛之间使用抑制消减杂交（SSH）来制备消减cDNA文库。由血细胞和造血器官制备单独的文库。在两个方向上进行减法，富集在寄生虫暴露的抗性和易感性样品之间差异表达的cDNA，以及攻击后抗性品系中上调或下调的cDNA。筛选了所得的8个文库，并通过逆转录酶PCR鉴定和确认了在抗性和易感性蜗牛品系的血细胞之间差异表达的8个基因，其中包括预计参与应激反应机制的两个转录本，用于调节参与细胞毒性杀灭寄生虫的破坏性氧化爆发途径：铁储存和免疫调节分子、铁蛋白和HtrA2（一种参与细胞应激反应的宁静蛋白酶）。抗性菌株中水平升高的转录本在扣除文库和未扣除对照中具有相同的表达模式；与易感蜗牛相比，暴露的抗性蜗牛的水平较高，与未暴露的抗性蜗牛相比，暴露的抗性蜗牛的水平下调。在重复暴露实验中独立证实了易感菌株中两个功能未知的转录物的差异表达。 (c) 2006 Elsevier B.V. 保留所有权利。
Biomphalaria glabrata is an intermediate snail host for Schistosoma mansoni, a medically important schistosome. In order to identify transcripts involved in snail-schistosome interactions, subtractive cDNA libraries were prepared, using suppression subtractive hybridization (SSH) between a parasite-exposed schistosome-resistant and a susceptible strain of B. glabrata, and also between schistosome-exposed and unexposed snails from the resistant snail line. Separate libraries were made from both haemocytes and the haemopoietic organ. Subtraction was performed in both directions enriching for cDNAs differentially expressed between parasite-exposed resistant and susceptible samples and up or down-regulated in the resistant line after challenge. The resulting eight libraries were screened and eight genes, differentially expressed between the haemocytes of resistant and susceptible snail strains, were identified and confirmed with reverse transcriptase PCR, including two transcripts expected to be involved in the stress response mechanism for regulating the damaging oxidative burst pathways involved in cytotoxic killing of the parasite: the iron-storage and immunoregulatory molecule, ferritin, and HtrA2, a serene protease involved in the cellular stress response. Transcripts with elevated levels in the resistant strain, had the same expression patterns in the subtracted libraries and unsubtracted controls; higher levels in exposed resistant snails compared to susceptible ones and down-regulated in exposed compared with unexposed resistant snails. Differential expression of two of the transcripts with no known function from the susceptible strain, was independently confirmed in a repeat exposure experiment. (c) 2006 Elsevier B.V. All rights reserved.