Sequence, Function and Expression of CCNC Gene in Ridgetail White Prawn Exopalaemon carinicauda
DAI Qin1, GAO Wei1, SHI Tingting1, CHEN Shuhan1, WANG Panpan1,2, LAI Xiaofang1,2, GAO Huan1,2,3, YAN Binlun1,2
1. Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, China; 2. The Jiangsu Provincial Infrastructure for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014, China; 3. Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Lianyungang 222005, China
Abstract：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.
戴琴, 高威, 史婷婷, 陈姝含, 王攀攀, 赖晓芳, 高焕, 阎斌伦. 脊尾白虾CCNC基因序列、功能与表达分析[J]. 水产科学, 2022, 41(6): 985-991.
DAI Qin, GAO Wei, SHI Tingting, CHEN Shuhan, WANG Panpan, LAI Xiaofang, GAO Huan, YAN Binlun. Sequence, Function and Expression of CCNC Gene in Ridgetail White Prawn Exopalaemon carinicauda. 水产科学, 2022, 41(6): 985-991.
[1]EVANS T, ROSENTHAL E T, YOUNGBLOM J, et al.Cyclin: a protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division[J]. Cell,1983,33(2):389-396. [2]万晶.细胞周期蛋白生物信息学分析[D].成都:西南交通大学,2012. [3]WOOD D J, ENDICOTT J A. Structural insights into the functional diversity of the CDK-cyclin family[J].Open Biology,2018,8(9):180112. [4]LIM S, KALDIS P. Cdks, yclins and CKIs: roles beyond cell cycle regulation[J].Development,2013,140(15):3079-3093. [5]JEEK JAN, SMETHURST D, STIEG D, et al. Cyclin C: the story of a non-cycling cyclin[J]. Biology,2019,8(3):1-23. [6]REN S J, ROLLINS B J. Cyclin C/cdk3 promotes Rb-dependent G0 exit[J].Cell,2004,117(2):239-251. [7]BOURBON H M. Comparative genomics supports a deep evolutionary origin for the large, four-module transcriptional mediator complex[J]. Nucleic Acids Research,2008,36(12):3993-4008. [8]JEZEK J, CHANG K T, JOSHI A M, et al. Mitochondrial translocation of cyclin C stimulates intrinsic apoptosis through Bax recruitment[J]. EMBO Reports,2019,20(9):e47425. [9]COOPER K F, KHAKHINA S, KIM S K, et al. Stress-induced nuclear-to-cytoplasmic translocation of cyclin C promotes mitochondrial fission in yeast[J]. Developmental Cell,2014,28(2):161-173. [10]RAJENDER P S, VASAVI M, VURUPUTURI U. Identification of novel selective antagonists for cyclin C by homology modeling and virtual screening[J]. International Journal of Biological Macromolecules,2011,48(2):292-300. [11]谢群,胡强,张瑞瑞,等.大面积重度烧伤患者CyclinB1、CyclinC及PCNA表达特征分析[J].中南医学科学杂志,2019,47(1):64-67. [12]JANPOOM S, PRASERTLUX S, RONGMUNG P, et al. Identification of a growth-associated single nucleotide polymorphism (SNP) in cyclin C of the giant tiger shrimp Penaeus monodon[J].Biochemical Genetics,2021,59(1):114-133. [13]梁俊平,李健,李吉涛,等.脊尾白虾全人工繁育技术初步研究[J].水产科学,2017,36(3):290-295. [14]栗治国,张成松,李富花,等.脊尾白虾的性腺发育及组织结构观察[J].水产学报,2014,38(3):362-370. [15]王绪峨.脊尾白虾的幼体发育[J].海洋科学,1991,15(1):12-14. [16]薛蓓,张培,李志辉,等.脊尾白虾GAPDH基因的克隆及其内参基因稳定性分析[J].中国水产科学,2017,24(5):1003-1012. [17]LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J]. Methods,2001,25(4):402-408 [18]MALUMBRES M. Cyclin-dependent kinases[J]. Genome Biology,2014,15:122. [19]STRICH R, COOPER K F. The dual role of cyclin C connects stress regulated gene expression to mitochondrial dynamics[J]. Microbial Cell,2014,1(10):318-324. [20]杨亚男,叶海辉,尚丽丽,等.刀额新对虾周期蛋白B基因的分子克隆及序列分析[J].厦门大学学报(自然科学版),2012,51(4):753-758. [21]贾复龙,孟宪亮,刘萍,等.三疣梭子蟹细胞周期蛋白H基因克隆及其在卵巢发育中的表达分析[J].中国海洋大学学报(自然科学版),2016,46(7):62-69. [22]郭松,傅明骏,赵超,等.斑节对虾细胞周期蛋白T基因的克隆和表达分析[J].南方水产科学,2016,12(1):59-66. [23]李伟杰,傅明骏,赵超,等.斑节对虾细胞周期蛋白Y基因克隆与原核表达[J].中国水产科学,2015,22(5):896-907. [24]韩坤煌.拟穴青蟹Sp-cycH基因的分子克隆与表达分析[J].宁德师范学院学报(自然科学版),2016,28(2):155-160. [25]董胜张.蝶蛹金小蜂卵黄发生与卵子发生的生理与分子基础[D].杭州:浙江大学,2007.
2022, Vol. 41 
