三角帆蚌基质蛋白基因cysrichin-F序列与功能分析  

Sequence and functional analysis of the matrix protein gene cysrichin-F in Hyriopsis cumingii

作　　者：梁晨晨 刘晓军 汪桂玲[1] LIANG Chenchen;LIU Xiaojun;WANG Guiling(Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Department of Biotechnology and Biomedicine,Yangtze Delta Region Institute of Tsinghua University,Jiaxing 314000,Zhejiang,China)

机构地区：[1]上海海洋大学农业农村部淡水水产种质资源重点实验室,上海201306 [2]浙江清华长三角研究院生物技术与医药研究所,浙江嘉兴314000

出　　处：《上海海洋大学学报》2025年第1期25-33,共9页Journal of Shanghai Ocean University

基　　金：国家自然科学基金(32072975)。

摘　　要：基质蛋白在调控无机矿物质形成过程中起着至关重要的作用,为了探究三角帆蚌(Hyriopsis cumingii)基质蛋白对贝壳生物矿化的影响,从三角帆蚌中分离获得一个新的基质蛋白基因cysrichin-F,并进一步利用原位杂交、破壳修复、RNA干扰等技术探究了该基因的生物学功能。结果显示,cysrichin-F序列全长为592 bp,开放阅读框区域(Open reading frame,ORF)为42~389 bp,共编码115个氨基酸。该基因编码的蛋白理论分子量为12.895 ku,理论等电点为8.65,为碱性蛋白,半胱氨酸富集-F(CYSRICHIN-F)的前23个氨基酸为信号肽。实时荧光定量PCR (qRT-PCR)检测发现,该基因在外套膜中的表达量相对其他组织更高。进一步通过原位杂交和破壳修复技术发现,cysrichin-F参与了贝壳角质层和棱柱层的形成过程,该基因被干扰后,贝壳棱柱层的结构将被破坏,最终导致贝壳棱柱层形状不规则。研究表明,cysrichin-F参与了贝壳角质层和棱柱层的生物矿化过程,研究结果有助于阐明三角帆蚌生物矿化的分子机制,为三角帆蚌基质蛋白在贝壳形成过程中发挥的作用提供一定的理论基础。Matrix proteins play a crucial role in regulating the formation of inorganic minerals.In order to investigate the effect of matrix proteins in Hyriopsis cumingi on shell biomineralization,this study isolated a new matrix protein gene cysrichin-F from H.cumingi,and further explored its biological function using techniques such as in situ hybridization,shell-breaking repair and RNA interference(RNAi).The results showed that the full length of cysrichin-F sequence was 592 bp,and the open reading frame(ORF)region was 42-389 bp,encoding a total of 115 amino acids.The protein encoded by this gene has a theoretical molecular weight of 12.895 ku and a theoretical isoelectric point of 8.65,making it an alkaline protein.The front 23 amino acids of CYSRICHIN-F are signal peptides.Quantitative Real-time PCR(qRT-PCR)detection revealed that the expression level of this gene in the mantle was relatively high compared to other tissues.Through in situ hybridization and shell-breaking repair techniques,it was found that cysrichin-F was involved in the formation of the cuticle and prismatic layers of shells.After interference with this gene,the structure of the shell prismatic layer would be disrupted,ultimately resulting in an irregular shape of the shell prismatic layer.This study showed that cysrichin-F was involved in the biomineralization process of the cuticle and prism layers of shells.The results of the study can help to elucidate the molecular mechanism of the biomineralization in H.cumingi,and provide a theoretical basis for the role of matrix proteins in the formation of shells in H.cumingi.

关 键 词：三角帆蚌 基质蛋白 原位杂交 破壳修复 RNA干扰 

分 类 号：S917[农业科学—水产科学]