Screening and expression of cyclins gene in Hyriopsis cumingii

doi:10.3969/j.issn.1004-1524.2021.11.06

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (11): 2041-2050.DOI: 10.3969/j.issn.1004-1524.2021.11.06

• Animal Science • Previous Articles Next Articles

Screening and expression of cyclins gene in Hyriopsis cumingii

FENG Shangle^a(), LI Xuenan^a, CHEN Yige^a, LIU Ruiqi^a, BAI Zhiyi^a^,^b^,^c, LI Wenjuan^a^,^b^,^c^,^*()

a. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China
b. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China
c. National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China

Received:2021-01-16 Online:2021-11-25 Published:2021-11-26
Contact: LI Wenjuan

Abstract

Abstract:

Cyclins plays an important role in cell cycle entry and progression, they can modulate the catalytic activities of cyclin-dependent kinases (CDKs), cyclins can binding the catalytic subunits of CDKs to regulate cell cycle.To select cyclin genes in Hyriopsis cumingii, explore the function of cyclins, total RNA from mixed sample of mantle, gonad, visceral mass and hemolymph were isolated and sequenced by Illumina HiSeq 2000.Based on the sequencing results, select the cyclin A, cyclin D2, cyclin E genes, through cloning their conserved sequence to verify cyclins genes in H. cumingii. Amino acid sequence similarity analysis showed that cyclin A, cyclin D2 and cyclin E of H. cumingii have high sequence identity with Crassostrea virginica and Crassostrea gigas. At the transcriptional level, quantitative real-time PCR (qRT-PCR) was used to detect expression of cyclin genes in different tissues. The result revealed that all cyclin genes were expressed in tested tissues (adductor, foot, visceral mass, gill, heart, mantle, gonad, blood) with variable levels, expression levels of cyclin A and cyclin E genes in gonad were significantly (P<0.05) higher than those in other tissues, expression level of cyclin D2 gene in gills was significantly(P<0.05) higher than those in other tissues.Between different genders in the same tissue, there were significant(P<0.05) gender differences in the relative expression of cyclin A gene in gonad, adductor, heart and blood.In gonads and heart,there were significant(P<0.05) gender differences in cyclin D2 and cyclin E expression.Cyclin A and cyclin E genes were significantly(P<0.05)higher expressed in female hearts and gonads than those in males,indicated that cyclin A and cyclin E genes play an important role in gonad development.The high expression of cyclin D2 in gill suggested that it might be associated with gill damage repair. In this study, cyclin A, cyclin D2 and cyclin E genes and their functions were preliminarily studied, which laid a molecular basis for further regulation of the cell growth in vitro.

Key words: transcriptome, Hyriopsis cumingii, cyclins, tissue expression profile

CLC Number:

S634.1

FENG Shangle, LI Xuenan, CHEN Yige, LIU Ruiqi, BAI Zhiyi, LI Wenjuan. Screening and expression of cyclins gene in Hyriopsis cumingii[J]. Acta Agriculturae Zhejiangensis, 2021, 33(11): 2041-2050.

Figures/Tables 11

Table 1 Primers used in the experiment

引物Primer	上游引物(5'-3')Forward primer(5'-3')	下游引物(5'-3')Reverse primer(5'-3')
Cyclin A	AGCAATGACACTTGGGATG	TTGTAGAGGTGGCAGAGG
Cyclin D2	TGTGTGTAGAGGCCGAATCA	TCCCTAGTGAAAGCCGACAC
Cyclin E	TATCCCCAATTCTGAGCATGGA	TATCACCTGGGCCTGAATGTC
Cyclin A-Q Cyclin D2-Q Cyclin E-Q EF1α	CACATCAGGGGGATAGAT GATAGCAGCAGGGAGTGT GTCCCGTCTGAGGATAGC GGAACTTCCCAGGCAGACTGTGC	AGAGGTGGCAGAGGAATA TTTGTGGATTCCGTTTCG CTCTGTGGAGGTGTAAGC TCAAAACGGGCCGCAGAGAAT

Table 2 Screening of cyclin related genes in transcriptome library of Hyriopsis cumingii

基因ID Gene ID	基因 Gene	长度 Length/bp
comp121579_c0_seq1	cyclin A	1 929
comp136327_c0_seq1	cyclin A	3 881
MblContig8891	G2/mitotic-specific cyclin B	3 641
MblContig41239 MblContig65698 MblContig31421 MblContig39154 MblContig1030 comp77562_c0_seq1 MblContig9313	cyclin B3 cyclin B G1/S-specific cyclin D2 G1/S-specific cyclin-D2 G1/S-specific cyclin E cyclin G1 cyclin H	835 1 935 1 971 4 358 1 019 659 2 006

Fig.1 Predicted amino acid sequence of cyclin A gene in Hyriopsis cumingii Black frame was the cyclin box.

Fig.2 Amino acid similarity alignment of cyclin A among Hyriopsis cumingii and other species GenBank accession numbers of cyclin A: Dreissena polymorpha, AAC35953.1;Crassostrea virginica, XP_022320685.1; Crassostrea gigas, XP_034299932.1;Mizuhopecten yessoensis, XP_021364053.1;Penaeus monodon, ACH72071.1;Xenopus laevis, NP_001081579.1;Danio rerio, AAH68323.2;Acanthaster planci, XP_022085019.1.

Fig.3 Predicted amino acid sequence of cyclin D2 gene in Hyriopsis cumingii Black frame was the cyclin box.

Fig.4 Amino acid sequence similarity alignment of cyclin D2 among Hyriopsis cumingii and other species GenBank accession numbers of cyclin D2: Dreissena polymorpha, AAM44813.1;Crassostrea virginica, XP_022292709.1;Crassostrea gigas, XP_011430967.2;Mizuhopecten yessoensis, XP_021360132.1;Xenopus laevis, CAA61665.1;Danio rerio, NP_001082914.1;Acanthaster planci, XP_022112210.1.

Fig.5 Predicted amino acid sequence of cyclin E gene in Hyriopsis cumingii

Fig.6 Amino acid sequence similarity alignment of cyclin E among Hyriopsis cumingii and other species GenBank accession numbers of cyclin E: Crassostrea virginica, XP_022336414.1;Crassostrea gigas, XP_011433150.1;Mizuhopecten yessoensis, OWF54564.1;Penaeus monodon, AGW23550.1;Xenopus laevis, CAA78370.1;Danio rerio, CAA58574.1; Acanthaster planci, XP_022092193.1.

Fig.7 Relative expression level of cyclin A Values without the same letters were significantly different (P<0.05);* indicated P<0.05, ** indicated P<0.01. The same as below.

Fig.8 Relative expression level ofcyclin D2

Fig.9 Relative expression level of cyclin E

References 27

[1]	李家乐, 王德芬, 白志毅, 等. 中国淡水珍珠养殖产业发展报告[J]. 中国水产, 2019(3):23-29.
	LI J L, WANG D F, BAI Z Y, et al. Report on the development of freshwater pearl culture industry in China[J]. China Fisheries, 2019(3):23-29.
[2]	李倩, 施志仪, 李文娟, 等. 三角帆蚌外套膜细胞体外培养优化及体内植入培养对细胞生长的影响[J]. 中国水产科学, 2014, 21(2):225-234.
	LI Q, SHI Z Y, LI W J, et al. Impact of in vitro optimization and in vivo implantation culture on the growth of Hyriopsis cumingii mantle cells[J]. Journal of Fishery Sciences of China, 2014, 21(2):225-234.(in Chinese with English abstract)
[3]	施志仪, 李巍, 李松荣, 等. 三角帆蚌外套膜细胞培养与组织培养的比较[J]. 上海水产大学学报, 2002, 11(1):27-30.
	SHI Z Y, LI W, LI S R, et al. Comparision of the tissue culture and cell culture from the mantle epithelium of Hyriopsis cumingii[J]. Journal of Shanghai Fisheries University, 2002, 11(1):27-30.(in Chinese with English abstract)
[4]	HARTWELL L H, KASTAN M B. Cell cycle control and cancer[J]. Science, 1994, 266(5192):1821-1828. DOI URL
[5]	NURSE P. A long twentieth century of the cell cycle and beyond[J]. Cell, 2000, 100(1):71-78. DOI URL
[6]	MARTÍNEZ-ALONSO D, MALUMBRES M. Mammalian cell cycle cyclins[J]. Seminars in Cell & Developmental Biology, 2020, 107:28-35.
[7]	HUANG H Y, LI W X, HUANG J R, et al. Two transcripts and the expression profiles of cyclin A in ovary of the mud crab, Scylla paramamosain[J]. Marine and Freshwater Behaviour and Physiology, 2013, 46(1):45-58. DOI URL
[8]	LI W X, HUANG H Y, HUANG J R, et al. Evidence for cyclin A and cyclin B genes in the ovary of the horseshoe crab, Tachypleus tridentatus: molecular cloning and expression profiles[J]. Marine and Freshwater Behaviour and Physiology, 2015, 48(3):163-176. DOI URL
[9]	VISUDTIPHOLE V, KLINBUNGA S, KIRTIKARA K. Molecular characterization and expression profiles of cyclin A and cyclin B during ovarian development of the giant tiger shrimp Penaeus monodon[J]. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2009, 152(4):535-543. DOI URL
[10]	HAN K H, DAI Y B, ZOU Z H, et al. Molecular characterization and expression profiles of cdc2 and cyclin B during oogenesis and spermatogenesis in green mud crab (Scylla paramamosain)[J]. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2012, 163(3/4):292-302. DOI URL
[11]	VORONINA E, MARZLUFF W F, WESSEL G M. Cyclin B synjournal is required for sea urchin oocyte maturation[J]. Developmental Biology, 2003, 256(2):258-275. DOI URL
[12]	ZHAO C, FU M J, ZHOU F L, et al. Characterization and expression analysis of a cyclin B gene from black tiger shrimp (Penaeus monodon)[J]. Genetics and Molecular Research, 2015, 14(4):13380-13390. DOI URL
[13]	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.
	YANG Y N, YE H H, SHANG L L, et al. Molecular cloning and analysis of cyclin B from shrimp Metapenaeus ensis[J]. Journal of Xiamen University (Natural Science), 2012, 51(4):753-758.(in Chinese with English abstract)
[14]	蔡明夷, 周鹏, 韩坤煌, 等. 大黄鱼cyclin B1和cdc2 cDNA序列特征及组织表达分析[J]. 厦门大学学报(自然科学版), 2014, 53(1):132-141.
	CAI M Y, ZHOU P, HAN K H, et al. Characterization and tissue expression profiles of cyclin B1 and cdc2 in large yellow croaker(Larimichthys crocea)[J]. Journal of Xiamen University (Natural Science), 2014, 53(1):132-141.(in Chinese with English abstract)
[15]	MOORE J C, SUMEREL J L, SCHNACKENBERG B J, et al. Cyclin D and cdk4 are required for normal development beyond the blastula stage in sea urchin embryos[J]. Molecular and Cellular Biology, 2002, 22(13):4863-4875. DOI URL
[16]	KUROKAWA D, AKASAKA K, IVIITSUNAGA-NAKATSUBO K, et al. Cloning of cyclin E cDNA of the sea urchin, Hernicentrotus pulcherrimus[J]. Zoological Science, 1997, 14(5):791-794. DOI URL
[17]	王桐, 李莉, 阙华勇, 等. 长牡蛎细胞周期调控关键基因cyclin B3的克隆及其在性腺发育中的作用[J]. 海洋科学, 2011, 35(12):1-9.
	WANG T, LI L, QUE H Y, et al. Molecular cloning and characterization of the key regulator of cell cycle cyclin B3 in Pacific Oyster (Crassostrea gigas), and its role in gonad development[J]. Marine Sciences, 2011, 35(12):1-9.(in Chinese with English abstract)
[18]	刘佳敏, 李文娟, 施志仪, 等. 三角帆蚌cyclin B基因克隆及功能[J]. 上海海洋大学学报, 2020, 29(4):496-505.
	LIU J M, LI W J, SHI Z Y, et al. Expression and function of cyclin B gene in Hyriopsis cumingii[J]. Journal of Shanghai Ocean University, 2020, 29(4):496-505.(in Chinese with English abstract)
[19]	吴丛迪, 陈亚, 汪桂玲, 等. M、F型ATP8基因在雌雄三角帆蚌不同发育时期的表达差异研究[J]. 基因组学与应用生物学, 2018, 37(10):4289-4296.
	WU C D, CHEN Y, WANG G L, et al. The expression of M and F-type ATP8 genes in different ages freshwater mussel Hyriopsis cumingii[J]. Genomics and Applied Biology, 2018, 37(10):4289-4296.(in Chinese with English abstract)
[20]	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. DOI URL
[21]	COQUERET O. Linking cyclins to transcriptional control[J]. Gene, 2002, 299(1/2):35-55. DOI URL
[22]	周开梅, 郭瑞珍. 细胞周期蛋白在恶性肿瘤中的表达[J]. 医学综述, 2010, 16(4):533-536.
	ZHOU K M, GUO R Z. Expression of cyclins in malignant tumors[J]. Medical Recapitulate, 2010, 16(4):533-536.(in Chinese with English abstract)
[23]	FAIVRE J, FRANK-VAILLANT M, POULHE R, et al. Membrane-anchored cyclin A2 triggers Cdc2 activation in Xenopus oocyte[J]. FEBS Letters, 2001, 506(3):243-248. DOI URL
[24]	赵超, 傅明骏, 江世贵, 等. 斑节对虾细胞周期蛋白E基因的克隆与表达分析[J]. 中国水产科学, 2014, 21(3):464-473.
	ZHAO C, FU M J, JIANG S G, et al. Molecular cloning and expression analysis of the cyclin E gene from black tiger shrimp Penaeus monodon[J]. Journal of Fishery Sciences of China, 2014, 21(3):464-473.(in Chinese with English abstract)
[25]	DARZYNKIEWICZ Z, GONG J P, JUAN G, et al. Cytometry of cyclin proteins[J]. Cytometry, 1996, 25(1):1-13. DOI URL
[26]	卢梦玲, 闫超, 赖多, 等. Cyclin D1与细胞周期调控[J]. 生物技术通报, 2011(10):55-59.
	LU M L, YAN C, LAI D, et al. Cyclin D1 and cell cycle regulation[J]. Biotechnology Bulletin, 2011(10):55-59.(in Chinese with English abstract)
[27]	骆大鹏, 余祥勇, 王梅芳, 等. 三角帆蚌鳃的组织学研究[J]. 广东海洋大学学报, 2008, 28(3):6-11.
	LUO D P, YU X Y, WANG M F, et al. Histological study on the inner and outer gill of Hyriopsis cumingii[J]. Journal of Guangdong Ocean University, 2008, 28(3):6-11.(in Chinese with English abstract)

Screening and expression of cyclins gene in Hyriopsis cumingii

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 11

References 27

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	MA Jie, QU Wen, CHEN Chunyan, WANG Lei, MA Jun, LIU Zhenshan, MA Wei, ZHOU Ping, HE Yuankuan, SUN Bo. Development of SSR markers based on transcriptome sequencing and genetic diversity analysis of Nainaiqingcai leaf mustard [J]. Acta Agriculturae Zhejiangensis, 2021, 33(9): 1640-1649.
[2]	HUANG Changbing, CHENG Peilei, YANG Shaozong, ZHANG Huanchao, JIANG Zhengzhi, JIN Limin. Transcriptome analysis of Hemerocallis fulva under low temperature stress [J]. Acta Agriculturae Zhejiangensis, 2021, 33(8): 1445-1460.
[3]	JIANG Zhifang, HAN Yidie, LOU Panpan, GUO Hong, FENG Shangguo, SHEN Chenjia, WANG Huizhong1. Identification and expression analysis of cytochrome P450 family genes from Physalis angulata L. [J]. Acta Agriculturae Zhejiangensis, 2021, 33(11): 2009-2016.
[4]	YIN Minghua, CAO Qing, CHEN Hong, DENG Siyu, DENG Yanmei. Transcriptome analysis of red bud taro and green stem taro in Yanshan, Jiangxi Province [J]. , 2020, 32(9): 1533-1543.
[5]	LIU Xinyu, TIAN Jie. Analysis of simple sequence repeats in transcriptome of garlic (Allium sativum L.) and development of molecular markers [J]. , 2020, 32(9): 1615-1625.
[6]	GE Jintao, WANG Jiangying, ZHAO Wenjing, SHAO Xiaobin, ZHU Pengbo, TANG Xueyan, SUN Mingwei, LIU Xingman. Transcriptome analysis on development of aerial root in grape of Weike [J]. , 2020, 32(9): 1645-1655.
[7]	ZHU Yu, LIU Yang. Transcriptome analysis on heat tolerance of Chilo suppressalis larvae [J]. , 2020, 32(5): 849-857.
[8]	ZHU Xiaolin, WEI Xiaohong, WANG Baoqiang, WANG Xian, ZHANG Mingjun. Transcriptome analysis of tomato under salt stress induced by c-GMP [J]. , 2020, 32(10): 1788-1797.
[9]	WANG Qi, CHEN Xiaojie, GU Shuangyue, ZHANG Xinyue, HANG Tianlu, DING Ting. Transcriptome profiling of maize resistance gene in response to DZSY21 induction [J]. , 2019, 31(3): 345-354.
[10]	WANG Hua, WANG Wangwei, WANG Dongliang, ZHANG Shihu, HU Xinfang, LU Shiyu, GONG Xuemei. De novo assembly and functional annotation of transcriptome data of Rhododendron pulchurum cv. Baifeng 4 leaf [J]. , 2018, 30(7): 1149-1159.
[11]	FENG Chen, TANG Haoru, JIANG Leiyu, SONG Xia, ZHANG Yunting, YE Yuntian, CHEN Qing, SUN Bo. Analysis of codon usage bias of specific genes in strawberry transcriptome under the red and blue light [J]. , 2017, 29(4): 566-574.
[12]	HONG Senrong, WU-XIA Junpeng, XU Wenhui, ZHAN Xuelin, XIE Ni’ni, JIANG Yan, WANG Jinhua, LING Fei, WU Lixia, WAN Lin. Correlation analysis of transcriptome, proteome and metabolome of Dioscorea bulbifera L. microtubers conserved in vitro at low temperature [J]. , 2017, 29(11): 1827-1834.
[13]	PEI Cuiming， ZHANG Zhenya， MA Jin*. Identification and analysis of salt stressresponsive transcription factor in leaf of southern type alfalfa [J]. , 2016, 28(4): 550-.
[14]	MA Jing 1， CHENG Tielong 1， 2， *， SUN Canyue 3， DENG Nan 1， SHI Shengqing 1， JIANG Zeping 1. Characterization of transcriptome reveals pathway of flavonoids in Ephedra sinica Stapf [J]. , 2016, 28(4): 609-.
[15]	YANG Min, HUANG Qiao, CHEN Liang-biao. Construction of type Ⅲ antifreeze protein eukaryotic expression plasmid and expression in zebrafish cell line [J]. , 2016, 28(11): 1862-1866.