三角帆蚌EF-hand基因的表达研究  

Study on the expression of EF-hand gene in Hyriopsis cumingii

作　　者：祁晓翔 李文娟[1] 施志仪[1,2] 尚朝 周子睿 尚攀[1] 

机构地区：[1]上海海洋大学农业部水产种质资源与养殖生态重点开放实验室,上海201306 [2]上海海洋大学上海市高校水产养殖学研究院,上海201306

出　　处：《浙江农业学报》2015年第12期2078-2085,共8页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金(31201991);教育部博士点基金(20123104120003);上海市优青项目(ssc11004)

摘　　要：淡水贝类包含EF-hand结构域的蛋白质对Ca^(2+)代谢具有重要的调控作用,加强EF-hand蛋白的研究对培育大型优质珍珠具有十分重要的意义。通过三角帆蚌转录组筛选到7个EF-hand基因,并进行生物信息学分析;通过数字基因表达谱(DGE)分析了这7个基因在不同育珠时间两个育珠组织(外套膜和内脏团)中的表达趋势;通过Real-time Q-PCR技术对EF-hand基因进行了表达研究。结果表明,三角帆蚌7个EFhand基因都具有经典保守的EF-hand模块。进化树分析表明,EFCB1和EFCB6进化关系最近,EFCB7和N-EFCB1进化关系较近,EFCBD2和RASEFCB进化关系较近。DGE表达趋势分析表明三角帆蚌EF-hand基因在外套膜和内脏团组织不同育珠时期表达变化趋势差异较大,说明这7个基因在珍珠形成过程中的功能可能各不相同。荧光定量PCR结果表明,EF-hand基因在插核后不同时期的外套膜和内脏团育珠部位组织中表达与DGE表达趋势分析基本一致。EFCB1在外套膜插核后20 d表达水平最高(P<0.05),EFCB6和EFCB7在内脏团插核后20 d表达水平最高(P<0.05),表明这3个基因在珍珠质最初分泌形成过程中可能起着重要的作用,为进一步探索该类蛋白调控钙代谢及在珍珠形成过程中的作用奠定了基础。EF-hand domain-containing protein of pearl oyster played an important role in the regulation of calcium metabolism in pearl formation. Strengthening the study of the EF-hand protein was very important to cultivate large high-quality pearls. In this study, seven EF-hand genes were screened from transcriptome of Hyriopsis cumingii, then bioinformatics analysis of them was performed. Expression trends and levels of these seven EF-hand genes were analyzed at different periods in the mantle and visceral mass after inserted nucleus through the digital gene expression profiling （DGE） and real-time quantitative PCR. The results showed that these seven EF-hand genes had classic conservative EF-hand module. Phylogenetic analysis showed that EFCB1 and EFCB6, EFCITI and N-EFCB1, EF- CBD2 and RASEFCB had closer representation than the others. DGE analysis showed that EF-hand genes at differentculturing period had different expression levels in mantle and visceral mass organizations of Hyriopsis cumingii, sug- gesting that these seven genes had different functions during the formation of pearl. Fluorescence quantitative PCR re- suits of these seven genes in mantle and visceral mass at different periods after inserting nucleus showed the same trend as DGE analysis. EFCB1 showed the highest expression level in mantle at 20 days after nucleus inserting （P 〈 0. 05）, EFCB6 and EFCB7 showed the highest expression level in visceral mass at 20 days after nucleus inserting （P 〈 0. 05 ）, suggesting that these 3 genes might play important roles in the initial phase of nacre secretion forming process. This study provided useful information for further study on the regulation of calcium metabolism and pearl formation.

关 键 词：三角帆蚌 EF-hand基因 外套膜 内脏团 有核珍珠 

分 类 号：S966.23[农业科学—水产养殖] Q786[农业科学—水产科学]