| 泥蚶Smad1/5基因cDNA全全长克隆及时空表达特征分析 |
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| 引用本文： | 钱雪骏,董迎辉,姚韩韩,林志华.泥蚶Smad1/5基因cDNA全全长克隆及时空表达特征分析[J].水产学报,2015,39(9):1302-1312. |
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| 作者姓名： | 钱雪骏 董迎辉 姚韩韩 林志华 |
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| 作者单位： | 上海海洋大学水产与生命学院；浙江省水产种质资源高效利用技术研究重点实验室,浙江万里学院生物与环境学院,浙江万里学院生物与环境学院,浙江万里学院生物与环境学院 |
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| 基金项目： | 国家现代贝类产业技术体系项目（CARS-48）；浙江省自然科学基金重点项目（LZ12C19001）；浙江省重大科技专项（2012C12907-4）；上海海洋大学研究生科研基金项目（A1-0209-14-0900-68） |
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| 摘 要： | 为了研究Smad1/5基因在泥蚶生长、发育过程中的调控作用，本文利用SMART RACE方法克隆得到泥蚶Smad1/5基因（Tg-Smad1/5）的cDNA全长序列，该序列全长2 424 bp，开放阅读框有1 386 bp，编码462个氨基酸。Tg-Smad1/5蛋白与合浦珠母贝Smad5、太平洋牡蛎Smad5和大西洋舟螺Smad1的同源性分别达到了92.3%，91.2%和80.4%，与脊椎动物的同源性都在70%以上；该蛋白包含MH1和MH2区两个较为保守的结构域，此结构与高等动物Smad1和Smad5蛋白极为相似，表明该基因在物种进化过程中比较保守。利用qRT-PCR技术，研究了Smad1/5基因在泥蚶6个组织和9个发育时期中的表达情况，结果显示，Tg-Smad1/5基因在泥蚶成贝6个组织中均有表达，而在斧足中的表达量最高，极显著地高于其他组织；在各发育时期中，Tg-Smad1/5表达量随发育进程呈逐渐升高的趋势，在眼点幼虫期达到最高，极显著高于其他时期，而在变态至稚贝时，表达量又极显著地下降。研究结果表明，Tg-Smad1/5具有类似于高等动物的分子结构，并在泥蚶不同组织、不同发育时期表达有所差异，这为进一步研究该基因在贝类中的功能和作用机制奠定了重要基础。
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| 关 键 词： | 泥蚶 Smad1/5 基因克隆 时空表达 |
| 收稿时间： | 2015/2/14 0:00:00 |
| 修稿时间： | 6/1/2015 12:00:00 AM |
Cloning and spatiotemporal expression analysis of Smad1/5 gene in the blood clam Tegillarca granosa |
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| QIAN Xuejun,DONG Yinghui,YAO Hanhan and LIN Zhihua.Cloning and spatiotemporal expression analysis of Smad1/5 gene in the blood clam Tegillarca granosa[J].Journal of Fisheries of China,2015,39(9):1302-1312. |
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| Authors: | QIAN Xuejun DONG Yinghui YAO Hanhan and LIN Zhihua |
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| Institution: | College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China;Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China,Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China,Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China and Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China |
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| Abstract: | In order to explore the function of Smad1/5 during the process of the growth and development in Tegillarca granosa, Tg-Smad1/5 was cloned by SMART RACE techniques. The full length cDNA of Tg-Smad1/5 was 2 424 bp, and its complete ORF (Open Reading Frame) was 1 386 bp encoding 462 amino acids. The homologous similarity between the clam and Pinctada fucata Smad5, Crassostrea gigas Smad5, Crepidula fornicate Smad1 was 92.3%, 91.2% and 80.4%, respectively. It shared more than 70% similarity with vertebrates. There were two conserved domains of Smads family, MH1 and MH2, which was especially similar to the vertebrata. All above suggested that, Smad1/5 is relatively conserved in the evolution, and has a similar function in various species. The results of 6 tissue-specific expression by real time PCR showed that Tg-Smad1/5 gene expressed in all tissues, and the expression of foot showed significantly higher than other tissues. The results of relative expression in 9 development stages revealed that the expression of Tg-Smad1/5 gene showed the highest expression in the eyebot larvae stage, which was significantly higher than other stages. And the expression decreased significantly from eyebot larvae stage to juvenile clams. The results showed that Tg-Smad1/5 had the similar molecular structure as the vertebrata, and the relative expression revealed differences in different tissues and development stages, which laid the theoretical foundation for further study on function and mechanisms of Smad1/5 in molluscs. |
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| Keywords: | Tegillarca granosa Smad1/5 gene cloning spatiotemporal expression |
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