文章摘要
李杰,许国绿,沈和定,顾冰宁,杨铁柱.石磺钙调蛋白Os-IP3ROs-RyR基因的克隆、相对表达量及进化关系[J].水产学报,2018,42(12):1857~1868
石磺钙调蛋白Os-IP3ROs-RyR基因的克隆、相对表达量及进化关系
Clone and relative expression of Os-IP3R and Os-RyR to discuss the evolutionary relationship in Onchidiidae
投稿时间:2018-01-06  修订日期:2018-03-18
DOI:10.11964/jfc.20170610881
中文关键词: 石磺  Os-RyR  Os-IP3R  qRT-PCR  表达量百分比
英文关键词: Onchidiidae  Os-RyR  Os-IP3R  qRT-PCR  percentage of gene expression
基金项目:国家自然科学基金(41276157);上海高校水产学一流学科建设项目
作者单位E-mail
李杰 上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室, 上海 201306  
许国绿 上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室, 上海 201306  
沈和定 上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室, 上海 201306 hdshen@shou.edu.cn 
顾冰宁 上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室, 上海 201306  
杨铁柱 上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室, 上海 201306  
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中文摘要:
      为明确石磺钙通道蛋白1,4,5-三磷酸肌醇受体(IP3R)基因和蓝尼碱受体(RyR)基因的序列和结构信息,初步研究不同石磺中的Onchidium struma-IP3R/Onchidium struma-RyROs-IP3R/Os-RyR)基因表达量百分比与石磺系统进化的相关性,实验在瘤背石磺表皮转录组数据库的基础上,克隆得到2条钙通道蛋白基因Os-IP3ROs-RyR,利用生物信息学技术对该基因及所编码蛋白的结构特征进行分析,并通过qRT-PCR技术分析2个基因在瘤背石磺、平疣桑椹石磺和紫色疣石磺各组织中的表达情况。结果显示,Os-IP3R核酸序列为4 574 bp,包括2 808 bp的开放阅读框,共编码935个氨基酸,预测编码的蛋白有6个跨膜区;Os-RyR核酸序列为1 253 bp,包括1 131 bp的开放阅读框,共编码376个氨基酸,预测编码的蛋白有3个跨膜区。将瘤背石磺IP3R和RyR的氨基酸序列进行比对,发现位于钙离子通道区的G*R*GGG*GD序列处高度保守;发现3种石磺Os-IP3R/Os-RyR 的相对表达百分比的高低顺序与各石磺从陆地到浅海的梯度分布趋势相一致,依次为瘤背石磺>平疣桑椹石磺>紫色疣石磺;石磺的陆栖性越强,则Os-IP3R/Os-RyR 的相对表达百分比越高。不同种石磺的Os-IP3R/Os-RyR表达量百分比的研究能为分析海洋无脊椎动物由海洋向陆地进化学说提供新的分子生物学线索。
英文摘要:
      In this study, full-length cDNA of Os-IP3R and Os-RyR in Onchidium struma was cloned by transcriptome data mining and rapid amplification cDNA ends (RACE) methods, and we analyzed the relationship between the percentages of Os-IP3R/Os-RyR expression and the evolution of 3 species from family Onchidiidae. The bioinformatic analysis and expression pattern of Os-IP3R and Os-RyR in different tissues from Onchidium struma, Platevindex mortoni and Peronia verruculata detected by Real-time quantitative PCR were performed. The results show that length cDNA of Os-IP3R consists of 4 574 base pairs (bp), and 2 808 bp open reading frame (ORF) which encodes 935 amino acids. The predicted moleculer of the deduced amino acid of IP3R has six transmembrane domains. The length cDNA of Os-RyR consists of 1 253 base pairs (bp), and 1 131 bp open reading frame (ORF) which encodes 376 amino acids. The predicted molecular of the deduced amino acid of RyR has three transmembrane domains. The IP3R and RyR amino acid sequences in O. struma were aligned and found to be highly conserved in the G*R*GGG*GD at region of calcium channel. Os-IP3R and Os-RyR were detected by qRT-PCR in different tissues of three kinds of Onchidiidae, and the percentages of Os-IP3R/Os-RyR expression from three species were consistent with their living environment (from high tide zone to shallow sea), followed by O. struma, P. mortoni and P. verruculata. The species of higher terrestrial ability with higher percentages of Os-IP3R/Os-RyR gene expression, illustrated percentage of gene expression associated with evolutionary degree. According to the comparison of percentage in Onchidiidae, we hope to provide a valuable clue for amphibian investigators, and to give a molecular biology clue for studying sea to land transition of marine invertebrate.
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