| 摘要: |
| C-型凝集素是一种重要的模式识别蛋白,本研究以文蛤为材料。利用PCR扩增和RACE技术首次获得了编码文蛤C-型凝集素的基因(Mm-CTL)cDNA序列,全长为1855bp,其开放阅读框为519bp,编码172个氨基酸。Smart软件预测Mm-CTL的糖识别结构域(CRD)从第34个氨基酸到第168个氨基酸。BLASTP序列相似性分析显示, Mm-CTL与大竹蛏氨基酸序列同源性最高,相似性为56.5%,与其它无脊椎动物的相似性为13.5%-23.9%。系统进化树分析与同源分析结果一致,其中Mm-CTL与双壳类C-型凝集素聚在一起,与大竹蛏亲缘性最近。环境胁迫实验显示,溶藻弧菌感染6h时Mm-CTL表达量明显上升,在12h时达到最高值,随后有所下降。盐度胁迫时Mm-CTL在盐度5时表达量相对较低,在盐度10-20中的表达量相对较高。温度胁迫时Mm-CTL在35℃时表达量相对最高, 10℃时表达量明显较低。研究结果表明,文蛤应对环境因子影响可通过调节Mm-CTL的表达来应答免疫反应。本研究将为贝类先天性免疫因子研究以及病害的防控奠定基础。 |
| 关键词: 文蛤 C-型凝集素 基因克隆 生物信息学 环境因子 |
| DOI：10.11693/hyhz20150100017 |
| 分类号: |
| 基金项目:教育部长江学者与创新团队项目,IRT0734号;浙江省海水养殖重点科技创新团队项目,2010R50025-08号;宁波市水产病害防控与安全科技创新团队项目,2013B82012号。 |
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| MOLECULAR CLONE AND EXPRESSION OF C-TYPE LECTIN IN MERETRIX MERETRIX |
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LI Meng, ZHOU Su-Ming, LIU Lu, PENG Di, WANG Guo-Liang
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Key Laboratory of Applied Marine Biotechnology, Ministry of Education, School of Marine Science, Ningbo University, Ningbo 315211, China
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| Abstract: |
| The C-type lectin is one of the most important pattern recognition receptors. In present study, a C-type lectin was cloned from Meretrix meretrix(Mm-CTL) for the first time using degenerated PCR and rapid amplification of complementary DNA ends(RACE) method. The full length of the Mm-CTL cDNA was 1855bp containing a 519-bp open reading frame(ORF) encoding a 172-amino-acid polypeptide. Prediction of protein domains revealed that the Mm-CTL contain a typical carbohydrate recognition domain(CRD) that ranged 34th to 168th amino acid. BLASTP analysis showed that the Mm-CTL shared 56.5% identity with the ortholog in Solen grandis, and 13.5%-23.9% with other invertebrate orthologs. Mm-CTL is distinctly clustered with mollusk as expected in phylogenetic analysis. After Vibrio alginolyticus challenge, the Mm-CTL mRNA was induced significantly in 6h post challenge, and its expression peaked in 12h. Moreover, the Mm-CTL expression reduced in salinity 5 compared to that in salinity 10-20. In a thermal stress, the Mm-CTL expression was down-regulated at 10℃, whilst increased at 35℃. Therefore, the expression of Mm-CTL might response to bacterial infections or environmental stress. The findings may help understanding innate immune factor in molluscs for disease prevention. |
| Key words: Meretrix meretrix C-type lectin gene clone bioinformatics environmental factors |
