广西师范大学学报（自然科学版） ›› 2021, Vol. 39 ›› Issue (1): 136-147.doi: 10.16088/j.issn.1001-6600.2020042602

• 研究论文 • 上一篇    下一篇

卵形鲳鲹Myostatin基因克隆及其在胚胎发育中的表达分析

罗洪林, 冯鹏霏, 余艳玲, 肖蕊, 潘传燕, 宋漫玲, 张永德*   

  1. 广西水产科学研究院广西水产遗传育种与健康养殖重点实验室, 广西 南宁 530021
  • 收稿日期:2020-04-26 修回日期:2020-05-25 出版日期:2021-01-25 发布日期:2021-03-27
  • 通讯作者: 张永德(1977—), 男, 山东潍坊人, 广西水产科学研究院副研究员。E-mail: yondar@126.com
  • 基金资助:
    广西创新驱动发展专项资金项目(桂科AA17204080-3, 桂科AA18242031-2); 广西水产遗传育种与健康水产养殖重点实验室系统性课题(19-A-01-05)

Molecular Cloning of the Myostatin Gene and Its Expression During Embryo Development of Trachinotus ovatus

LUO Honglin, FENG Pengfei, YU Yanling, XIAO Rui, PAN Chuanyan, SONG Manling, ZHANG Yongde*   

  1. Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture, Guangxi Academy of Fishery Sciences, Nanning Guangxi 530021, China
  • Received:2020-04-26 Revised:2020-05-25 Online:2021-01-25 Published:2021-03-27

PDF (PC)

528

摘要： 为研究肌生长抑制素(myostatin,MSTN)在卵形鲳鲹胚胎发育过程中的表达,对卵形鲳鲹MSTN基因CDS区进行克隆与分析,并对其14个胚胎发育阶段的表达量进行研究。结果显示:克隆获得MSTN基因1 322 bp序列。该序列编码376个氨基酸,相对分子质量为42 694.85 u,理论等电点pI为5.52,为带负电的蛋白。MSTN蛋白质不存在跨膜结构域,亚细胞定位主要于细胞外部,可能属于分泌蛋白质,易于表达和纯化。MSTN蛋白质无信号肽序列,预测含有17个磷酸化位点。对24种鱼类MSTN基因序列的同源分析发现:卵形鲳鲹MSTN与布氏鲳鲹同源性最高,与黄尾鰤同源性次之;3种鱼类均属鲹科Carangidae,而鲹科鱼类与锯盖鱼科Centropomidae鱼类遗传距离最小,具有较近的亲缘关系。MSTN基因在卵形鲳鲹从受精卵到晶体出现期的13个阶段均表达量极低或不表达,组间差异不显著(P>0.05),而在初孵仔鱼期表达量迅速升高,极显著高于前13个时期(P<0.01)。MSTN基因可能在卵形鲳鲹孵化后发挥重要作用,而在孵化前作用微弱。

关键词: 卵形鲳鲹, MSTN, 基因克隆, 胚胎组织, 生物信息学分析

Abstract: Myostatin (MSTN) is a negative regulator of skeletal muscle growth,and much interest has been given to its role in animal development in recent years. In this study,MSTN gene in Trachinotus ovatus was cloned and characterized,and the expression patterns of 14 embryonic development stages of T. ovatus were studied. The results showed that 1 322 bp of MSTN gene was cloned,encoding a 376 amino acid polypeptide. The relative molecular weight was 42 694.85 u,and the theoretical isoelectric point pI was 5.52,which was a negatively charged protein. MSTN protein had no transmembrane domain and subcellular localization was mainly outside the cell,which may be secretory protein and easy to express and purify. MSTN protein had no signal peptide sequence,and 17 phosphorylation sites were predicted. A phylogenetic analysis of 24 species of fish revealed that the MSTN of T. ovatus had the highest homology with T. blochii,followed by that of S. lalandi dorsalis,all of which belonged to Carangidae,while that of Carangidae and centropomidae had the lowest genetic distance and close relationship. MSTN gene was very low or not expressed in all 13 stages of T. ovatus from Fertilized egg to Formation of eye lens stage,with no significant difference between the groups (P>0.05). However,the expression level of MSTN gene increased rapidly in the hatching stage,which was significantly higher than that in the previous 13 stages (P<0.01). The MSTN gene may play an important role after the T. ovatus hatched,but it is weak before hatching.

Key words: Trachinotus ovatus, myostatin, gene cloning, embryonic development, gene expression

中图分类号: 

  • S965.331
[1] 熊向英,王贤丰,彭银辉,等. 健康和患病卵形鲳鲹肠道菌群结构的差异[J]. 水产学报,2019,43(5): 1317-1325.DOI: 10.11964/jfc.20180411227.
[2] 胡海滨,钱雪桥,解绶启,等. 中成鱼阶段卵形鲳鲹对饲料蛋白质和脂肪的适宜需求量[J]. 上海海洋大学学报,2019,28(4): 566-576. DOI: 10.12024/jsou.20181202459.
[3] 易新文,陈瑞爱,徐家华. 鸡肉粉替代鱼粉对卵形鲳鲹生长、饲料利用和抗氧化力的影响[J]. 中国海洋大学学报(自然科学版),2019,49(12): 17-24. DOI: 10.16441/j.cnki.hdxb.20180203.
[4] TAN X H,SUN Z Z,ZHOU C P,et al. Effects of dietary dandelion extract on intestinal morphology,antioxidant status,immune function and physical barrier function of juvenile golden pompano Trachinotus ovatus[J]. Fish and Shellfish Immunology,2018,73: 197-206. DOI: 10.1016/j.fsi.2017.12.020.
[5] THOMAS M,LANGLEY B,BERRY C,et al. Myostatin,a negative regulator of muscle growth,functions by inhibiting myoblast proliferation[J]. Journal of Biological Chemistry,2000,275(51): 40235-40243.DOI: 10.1074/jbc.M004356200.
[6] McPHERRON A C,LAWLER A M,LEE S J. Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member[J]. Nature,1997,387(6628): 83-90. DOI: 10.1038/387083a0.
[7] MOSHER D S,QUIGNON P,BUSTAMANTE C D,et al. A mutation in the myostatin gene increases muscle mass and enhances racing performance in heterozygote dogs[J]. PLoS Genetics,2007,3(5): e79. DOI: 10.1371/journal.pgen.0030079.
[8] MEDEIROS E F,PHELPS M P,FUENTES F D,et al. Overexpression of follistatin in trout stimulates increased muscling[J]. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology,2009,297(1):R235-R242.
[9] LIN J,ARNOLD H B,DELLA-FERA M A,et al. Myostatin knockout in mice increases myogenesis and decreases adipogenesis[J]. Biochemical and Biophysical Research Communications,2002,291(3): 701-706.
[10] LI Q,LI Y G,HUANG X D,et al. Molecular cloning and single nucleotide polymorphisms identification of myostatin cDNA in the pearl oyster,Pinctada fucata[J]. Journal of the World Aquaculture Society,2014,45(6): 638-651.
[11] CHIANG Y A,KINOSHITA M,MAEKAWA S,et al. TALENs: mediated gene disruption of myostatin produces a larger phenotype of medaka with an apparently compromised immune system[J]. Fish and Shellfish Immunology,2016,48: 212-220.
[12] GAO F,SUN B X,XING S Y,et al. The effect of leader peptide mutations on the biological function of bovine myostatin gene[J]. Gene,2014,540(2): 171-177.
[13] HILL E W,GU J J,EIVERS S S,et al. A sequence polymorphism in MSTN predicts sprinting ability and racing stamina in thoroughbred horses[J]. PLoS ONE,2010,5(1): e8645.
[14] 李景芬,采克俊,曹访,等. 反向、巢式、降落PCR技术克隆奥尼罗非鱼MSTN基因3'侧翼序列[J]. 江苏农业科学,2014,42(2): 28-30.
[15] 李兴美,范巍,张彬,等. 鲤鱼肌肉生长抑制素基因(MSTN)的克隆及其组织表达特征[J]. 水生生物学报,2007,31(5): 643-648.
[16] 钟其旺. 几种鲆鲽鱼类Myostatin基因的克隆及结构和表达分析[D]. 青岛: 中国海洋大学,2007.
[17] 李盈,额尔敦木图,刘图雅,等. 肌肉生长抑制素基因在不同动物中的研究进展[J]. 中国畜牧兽医,2013,40(8): 141-145.
[18] 彭扣,赵浩斌,彭英,等. 普通鲤与建鲤MSTN基因比较分析[J]. 南昌大学学报(理科版),2013,37(2): 191-197.
[19] 区又君,李加儿. 卵形鲳鲹的早期胚胎发育[J]. 中国水产科学,2005,12(6): 786-788.
[20] 张猛,陈勇,沈玉帮,等. 草鱼 MSTN-1 基因多态性及与早期生长性状和肌肉成分关联分析[J]. 水产学报,2016,40(4): 618-625.
[21] 张冬杰,刘娣,张旭,等. 利用CRISPR-Cas9系统定点突变猪MSTN基因的研究[J]. 畜牧兽医学报,2016,47(1): 207-212.
[22] 王发新. 兰州鲇GH和MSTN基因多态性及生长相关性研究[D]. 兰州: 甘肃农业大学,2016.
[23] KONDRASHOV F A, ROGOZIN I B, WOLF Y I, et al. Selection in the evolution of gene duplications[J]. Genome Biology,2002,3(2): research0008.1.
[24] ZHANG S Y,LI Y,SHAO J J,et al. Functional identification and characterization of IpMSTNa,a novel orthologous myostatin (MSTN) gene in channel catfish Ictalurus punctatus[J]. International Journal of Biological Macromolecules,2020,152: 1-10.
[25] 俞兆曦. 兰州鲇生长性状相关基因的分离、序列特征及组织特异性表达分析[D]. 兰州: 甘肃农业大学,2016.
[26] 易恒洁,李辉,杨胜林,等. 三穗鸭MyoG基因多态性与屠宰性状的相关性研究[J]. 中国畜牧杂志,2016,52(9): 5-10.
[27] 李婷. 黑鲷MSTNⅡ基因分子特征[D]. 宁波:宁波大学,2009.
[28] 孙冉冉. 黄条鰤肌肉生长因子的克隆及其表达规律的研究[D]. 大连: 大连海洋大学,2019.
[29] BETANCUR-R R,WILEY E O,ARRATIA G,et al. Phylogenetic classification of bony fishes[J]. BMC Evolutionary Biology,2017,17: 162.
[30] HELTERLINE D L I,GARIKIPATI D,STENKAMP D L,et al. Embryonic and tissue-specific regulation of myostatin-1 and -2 gene expression in zebrafish[J]. General and Comparative Endocrinology,2007,151(1): 90-97.
[31] VIANELLO S,BRAZZODURO L, DALLA VALLE L,et al. Myostatin expression during development and chronic stress in zebrafish (Danio rerio)[J]. Journal of Endocrinology,2003,176(1): 47-59.
[32] XU C,WU G,ZOHAR Y,et al. Analysis of myostatin gene structure,expression and function in zebrafish[J]. Journal of Experimental Biology,2003,206(22): 4067-4079.
[1] 易潭, 刘希良, 宾石玉, 王开卓, 吴萍, 褚武英, 陈韬. 鳜鱼Siniperca chuatsi红肌sMyHC1基因cDNA的克隆及其表达分析[J]. 广西师范大学学报（自然科学版）, 2014, 32(2): 175-180.
[2] 王开卓, 宾石玉, 李虹辉, 李玉珑, 刘知行, 张建社, 褚武英. 翘嘴鳜生长抑制素基因的克隆及其表达分析[J]. 广西师范大学学报（自然科学版）, 2013, 31(4): 109-114.
[3] 赵志常, 胡福初, 胡桂兵, 王惠聪, 杨转英, 苏纯兰, 李加强. 荔枝类黄酮糖基转移酶(UFGT)基因的克隆及其原核表达研究[J]. 广西师范大学学报（自然科学版）, 2011, 29(4): 104-110.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 杨丽婷, 刘学聪, 范鹏来, 周岐海. 中国非人灵长类声音通讯研究进展[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 1 -9 .
[2] 宾石玉, 廖芳, 杜雪松, 许艺兰, 王鑫, 武霞, 林勇. 罗非鱼耐寒性能研究进展[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 10 -16 .
[3] 刘静, 边迅. 直翅目昆虫线粒体基因组的特征及应用[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 17 -28 .
[4] 李兴康, 钟恩主, 崔春艳, 周佳, 李小平, 管振华. 西黑冠长臂猿滇西亚种鸣叫行为监测[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 29 -37 .
[5] 和鑫明, 夏万才, 巴桑, 龙晓斌, 赖建东, 杨婵, 王凡, 黎大勇. 滇金丝猴主雄应对配偶雌性数量的理毛策略[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 38 -44 .
[6] 付文, 任宝平, 林建忠, 栾科, 王朋程, 王宾, 黎大勇, 周岐海. 济源太行山猕猴种群数量和保护现状[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 45 -52 .
[7] 郑景金, 梁霁鹏, 张克处, 黄爱面, 陆倩, 李友邦, 黄中豪. 基于木本植物优势度的白头叶猴食物选择研究[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 53 -64 .
[8] 杨婵, 万雅琼, 黄小富, 袁旭东, 周洪艳, 方浩存, 黎大勇, 李佳琦. 基于红外相机技术的小麂(Muntiacus reevesi)活动节律[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 65 -70 .
[9] 李友邦, 农娟丽, 杨婉琳, 赵佳佳, 朱琪琪. 弄岗同域分布赤腹松鼠和红颊长吻松鼠活动节律研究[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 71 -78 .
[10] 程瑞, 何明先, 钟春英, 罗树毅, 武正军. 野生与人工繁育鳄蜥游泳能力比较[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 79 -86 .
版权所有 © 广西师范大学学报（自然科学版）编辑部
地址：广西桂林市三里店育才路15号　邮编：541004
电话：0773-5857325　E-mail: gxsdzkb@mailbox.gxnu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发