脊尾白虾CCNC基因序列、功能与表达分析  

Sequence, Function and Expression of CCNC Gene in Ridgetail White PrawnExopalaemon carinicauda

作　　者：戴琴 高威 史婷婷 陈姝含 王攀攀 赖晓芳 高焕 阎斌伦 DAI Qin;GAO Wei;SHI Tingting;CHEN Shuhan;WANG Panpan;LAI Xiaofang;GAO Huan;YAN Binlun(Jiangsu Key Laboratory of Marine Bioresources and Environment,Jiangsu Key Laboratory of Marine Biotechnology,Jiangsu Ocean University,Lianyungang 222005,China;The Jiangsu Provincial Infrastructure for Conservation and Utilization of Agricultural Germplasm,Nanjing 210014,China;Co-Innovation Center of Jiangsu Marine Bio-industry Technology,Lianyungang 222005,China)

机构地区：[1]江苏海洋大学,江苏省海洋生物资源与生态环境重点实验室,江苏省海洋生物技术重点实验室,江苏连云港222005 [2]江苏省农业种质资源保护与利用平台,江苏南京210014 [3]江苏省海洋生物产业技术协同创新中心,江苏连云港222005

出　　处：《水产科学》2022年第6期985-991,共7页Fisheries Science

基　　金：江苏省研究生科研与实践创新计划项目(KYCX18-2572);江苏省优势学科建设工程项目(PAPD);江苏省海洋生物资源与环境重点实验室开放课题基金资助项目(SH20191203).

摘　　要：脊尾白虾是我国重要的经济虾类,细胞周期蛋白C(CCNC)是细胞周期蛋白家族的一员,在细胞周期调控及转录调控等方面发挥着重要作用。为研究脊尾白虾CCNC(EcCCNC)基因在卵巢及幼体发育中的生物学功能,采用RACE技术克隆EcCCNC基因cDNA全长序列,实时荧光定量PCR技术研究EcCCNC基因在脊尾白虾不同组织、卵巢不同时期和幼体发育不同时期的相对表达量,并通过原核表达获得EcCCNC基因的体外重组蛋白。结果显示:EcCCNC基因cDNA全长1042 bp,包括82 bp的5′非编码区,156 bp的3′非编码区,804 bp的开放阅读框,编码267个氨基酸,预测蛋白质的分子质量为31.34 ku,理论等电点为5.50;聚类分析表明,脊尾白虾EcCCNC基因编码的氨基酸与凡纳滨对虾的聚为一支,氨基酸序列与凡纳滨对虾的相似性(92%)最高;EcCCNC基因在卵巢中特别是大生长期及溞状幼体Ⅱ期的相对表达量最高(P<0.05);EcCCNC基因的体外重组蛋白为45 ku,经WB验证含His标签。研究结果表明,EcCCNC基因在脊尾白虾卵巢及幼体发育过程中很可能扮演了十分重要的角色。本研究结果可为进一步探究脊尾白虾卵巢及幼体发育的机制提供参考。Ridgetail white prawn Exopalaemon carinicauda is an economically important shrimp species along the coast of China. Cyclin C (CCNC) as a member of the cyclin family plays an important role in cell cycle regulation and transcription regulation. In order to study the biological function of CCNC in ovarian and larval development of ridgetail white prawn, the cDNA sequence of EcCCNC gene was obtained using rapid amplification of cDNA ends method (RACE). The relative expression levels of EcCCNC gene were analyzed in different tissues and at different stages of ovarian and larval development by qRT-PCR. Recombinant protein was obtained by prokaryotic expression. The results showed that the full-length cDNA sequence of EcCCNC was 1042 bp, which contained a 5′-UTR of 82 bp, a 3′-UTR of 156 bp, and a 804 bp open reading frame (ORF) encoding 267 amino acids. The predicted molecular weight of the protein was 31.34kDa , and the theoretical isoelectric point was 5.50. Cluster analysis showed that EcCCNC and CCNC in Pacific white shrimp Litopenaeus vannamei were clustered. The amino acid sequence also had the highest homology with Pacific white shrimp (92%). EcCCNC had the highest relative expression levels in the ovary, especially the ovarian stage Ⅲ and the larval stage Ⅱ ( P <0.05). The recombinant protein of EcCCNC was 45 ku and had a His tag. The findings indicate that EcCCNC may play an important role in the development of ovary and larva of ridgetail white prawn, which provides a theoretical basis for further understanding of the mechanism of shrimp ovarian and larval development.

关 键 词：脊尾白虾 CCNC基因 性腺发育 幼体发育 

分 类 号：Q786[生物学—分子生物学]