文蛤亮氨酸氨肽酶LAP3的基因克隆及其在幼体不同发育时期、成体不同组织的表达分析

阮文斌; 董迎辉; 高晓艳; 刘晨珊; 林德海; 林志华

doi:10.3969/j.issn.0253-4193.2017.02.009

文蛤亮氨酸氨肽酶LAP3的基因克隆及其在幼体不同发育时期、成体不同组织的表达分析

doi: 10.3969/j.issn.0253-4193.2017.02.009

1.
浙江万里学院水产种质资源高效利用技术研究浙江省重点实验室, 浙江宁波 315100;上海海洋大学水产与生命学院, 上海 201306
2.
浙江万里学院水产种质资源高效利用技术研究浙江省重点实验室, 浙江宁波 315100

基金项目: 国家自然科学基金项目（31372527）；国家现代贝类产业技术体系项目（CARS-48）；国家水产种质资源平台运行服务项目（2015DKA30470）；苏北科技专项（BN2015059）。

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出版历程
- 收稿日期: 2016-04-27
- 修回日期: 2016-11-24

Gene cloning and expression analysis of leucine aminopeptidase LAP3 gene in different development stages of larvae and different tissues of adult of Meretrix meretrix

1.
Key Laboratory of Aquatic Germplasm Resources of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China;College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China
2.
Key Laboratory of Aquatic Germplasm Resources of Zhejiang, Zhejiang Wanli University, Ningbo 315100, China

摘要

摘要: 为探索贝类LAP3基因的结构和功能特征以及在生长发育过程中的作用，利用cDNA末端快速扩增（RACE）法获得文蛤LAP3（Mm-LAP3）基因的cDNA全长序列，并对其生物信息学、组织及发育时期表达特征进行了分析。结果显示，Mm-LAP3基因cDNA全长2 037 bp，ORF区1 254 bp，编码417个氨基酸；Mm-LAP3蛋白由Peptidase_M17超家族序列的N-端结构域和Peptidase_M17催化结构域组成。荧光实时定量PCR结果表明，Mm-LAP3基因在文蛤成体6个组织和幼体10个发育时期中均有表达，其中在斧足中的表达量显著高于其他组织（P<0.05），这与斧足蛋白含量丰富、代谢旺盛相关；在幼体10个发育时期中的表达量呈现逐渐升高的趋势，在D形幼虫时期达到最高，随后又有所降低，推测Mm-LAP3基因在早期发育时期参与了某些组织器官的形成。
- 文蛤 /
- 亮氨酸氨肽酶(LAP3) /
- 基因克隆 /
- 表达分析
Abstract: To explore the molecular structure and functional characteristics of LAP3 gene and its role in the process of growth and development, full length cDNA of LAP3 in Meretrix meretrix(Mm-LAP3)was cloned by rapid amplification of cDNA ends technique(RACE), then the bioinformatics and expression profiles in different adult tissues and developmental stages of larvae were analyzed. The results showed that the full length cDNA of Mm-LAP3 gene was 2 037 bp, containing a complete 1 254 bp ORF encoding 417 amino acids. There were two functional domains of Mm-LAP3 protein (Peptidase_M17 and N-terminal). The result of Q-RT-PCR indicated that Mm-LAP3 expressed in six tissues and ten developmental stages, but the expression of foot was significantly higher than other tissues. The expression of Mm-LAP3 gradually increased with the process of the development, and showed the highest in D-shaped larvae stage, and then the expression decreased, which suggested that Mm-LAP3 may be involved in the formation of certain organs in early developmental stages.
- Meretrix meretrix /
- leucine aminopeptidase /
- gene cloning /
- expression analysis

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