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鉴定日本血吸虫慢性感染钉螺中差异表达的基因。

Identification of differentially expressed genes in Oncomelania hupensis chronically infected with Schistosoma japonicum.

机构信息

School of Basic Medical Science, Wuhan University, Wuhan, Hubei Province 430071, China.

出版信息

Exp Parasitol. 2012 Apr;130(4):374-83. doi: 10.1016/j.exppara.2012.02.004. Epub 2012 Feb 10.

Abstract

Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum. The schistosome-snail interaction is biomedically important. To identify differentially expressed transcripts in O. hupensis chronically infected with S. japonicum, suppression subtractive hybridization (SSH) was used to construct a cDNA library in each direction for transcripts that are more abundantly enriched in head-foot part of the infected O. hupensis and for those that are more abundantly enriched in the uninfected, as head-foot part contains hemocytes and hemolymph which are associated with the snail internal defense system. After differential screening, 39 transcripts were identified, including nine and 30 transcripts enriched in infected and uninfected snails, respectively. Some of the transcripts have similar homology to available sequences in current databases, including transposase, caveolin-like protein, pancreatic trypsin inhibitor-like protein, prosaposin, glutathione s-transferase (GST), and several hypothetical proteins, while most of the transcripts do not match with any sequences in available databases. The identified transcripts were involved functionally in cell growth, metabolism, signal transduction, and immune responses. Two forward library transcripts and 11 reverse library transcripts were selected for real-time PCR, and 10 of them were confirmed to be consistent with the SSH results. It is intriguing to continue functional studies for some genes such as pancreatic trypsin inhibitor; a hypothetical protein (HS576367) related to calcium ion binding; GST; and several unknown proteins (HS576353 and HS576355). These identified differentially expressed genes may be key targets for understanding the molecular mechanism of co-existence during which the snail is unable to rid itself of the schistosome in chronic infection stage.

摘要

钉螺是日本血吸虫唯一的中间宿主。血吸虫 - 钉螺的相互作用在医学上具有重要意义。为了鉴定慢性日本血吸虫感染的钉螺中差异表达的转录本,采用抑制性消减杂交(SSH)技术构建了两个方向的 cDNA 文库,一个文库中包含在感染钉螺头足部富集的转录本,另一个文库中包含在未感染钉螺头足部富集的转录本。头足部包含与钉螺内部防御系统相关的血细胞和血淋巴。经过差异筛选,共鉴定出 39 个转录本,其中 9 个在感染钉螺中富集,30 个在未感染钉螺中富集。部分转录本与现有数据库中的序列具有相似的同源性,包括转座酶、窖蛋白样蛋白、胰蛋白酶抑制剂样蛋白、前涎素、谷胱甘肽 S-转移酶(GST)和几个假定蛋白,而大多数转录本与现有数据库中的任何序列都不匹配。鉴定出的转录本在细胞生长、代谢、信号转导和免疫反应等方面具有功能。选择了两个正向文库转录本和 11 个反向文库转录本进行实时 PCR,其中 10 个转录本与 SSH 结果一致。对一些基因(如胰蛋白酶抑制剂;与钙离子结合相关的假定蛋白(HS576367);GST;和几个未知蛋白(HS576353 和 HS576355))进行功能研究是很有趣的。这些鉴定出的差异表达基因可能是理解分子机制的关键靶点,该分子机制是在慢性感染阶段,钉螺无法摆脱血吸虫的共生共存。

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