广西师范大学学报(自然科学版) ›› 2018, Vol. 36 ›› Issue (2): 141-147.doi: 10.16088/j.issn.1001-6600.2018.02.020

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翘嘴鳜USP7基因cDNA的克隆及饥饿对其表达影响分析

张方亮1,邓小红2,朱晓彤1,徐蕾1,胡姜丽1,宾石玉1*   

  1. 1.广西师范大学生命科学学院,广西桂林 541006;
    2. 全州县水产技术推广站,广西桂林 541500
  • 收稿日期:2017-05-10 出版日期:2018-05-10 发布日期:2018-07-18
  • 通讯作者: 宾石玉(1963—), 男, 广西全州人, 广西师范大学教授, 博士。E-mail:binsy@gxnu.edu.cn
  • 基金资助:
    国家自然科学基金(31560718)

Cloning and Expression Analysis of Ubiquitin-Specific Protease 7 Gene under Starvation in Mandarin Fish (Siniperca chuatsi)

ZHANG Fangliang1,DENG Xiaohong2,ZHU Xiaotong1,XU Lei1,HU Jiangli1,BIN Shiyu1*   

  1. 1. College of Life Science, Guangxi Normal University, Guilin Guangxi 541006,China;
    2.Quanzhou Fisheries Technology Station, Guilin Guangxi 541500,China
  • Received:2017-05-10 Online:2018-05-10 Published:2018-07-18

摘要: 为探讨饥饿对翘嘴鳜肝脏USP7基因表达的影响,本文应用实时荧光定量PCR、3′-RACE和5′-RACE技术,从翘嘴鳜肝脏组织中克隆得到了USP7基因的全长cDNA序列(登录号:MF000683)为4 319 bp。生物信息学分析表明:翘嘴鳜USP7基因编码区全长为3 336 bp,共编码个1 111氨基酸,其中180个带负电荷,137个带正电荷。翘嘴鳜USP7在饥饿中的表达分析表明,肝脏USP7基因在不同饥饿时间的相对表达量呈现一个峰值变化,在饥饿4 d时出现显著的上升且到达最高值;在饥饿7 d时又逐渐下降,最终达到初始水平(P<0.05)。本文结果发现饥饿后肝脏中USP7表达量的变化可能与翘嘴鳜的生长和健康状况密切相关,为鳜鱼的生产养殖提供一定的理论依据。

关键词: 翘嘴鳜, 泛素特异性蛋白酶7基因(USP7), 肝脏, 表达分析

Abstract: To investigate the effect of starvation on the expression of USP7 gene in the liver of Mandarin fish, the full-length cDNA sequence of the USP7 gene was cloned from the liver tissue of Mandarin fish by 3′-RACE and 5′-RACE techniques (Accession No. MF000683) with 4 319 bp and its expression profile upon starvation was assayed by real-time quantitative PCR. Bioinformatics analysis showed that the full-length coding region of USP7 gene is 3 336 bp with 1 111 encoded amino acids. Among them, 180 and 137 ones are negatively and positively charged, respectively. The relative expression of USP7 gene in the liver at different starvation time showed a peak change, which had a significant increase and reached the highest value after a starvation duration of 4 days, then decreased gradually after a starvation duration of 7 days and finally reached the initial level (P<0.05). Our studies provided novel information about the USP7gene and its expression status upon starvation.

Key words: Mandarin fish(Siniperca chuatsi), USP7 gene, liver, expression analysis

中图分类号: 

  • Q959.483
[1] ZHU X, CHEN D, HU Y, et al. The microRNA signature in response to nutrient restriction and refeeding in skeletal muscle of Chinese Perch (Siniperca chuatsi)[J]. Marine Biotechnology, 2015, 17(2): 180-189.
[2] 谢小军, 邓利, 张波. 饥饿对鱼类生理生态学影响的研究进展[J]. 水生生物学报, 1998, 22(2): 181-188.
[3] 孙红梅, 黄权, 丛波. 饥饿对黄颡鱼血液中几种免疫相关因子的影响[J]. 大连海洋大学学报, 2006, 21(4): 307-310.
[4] YAMAMURA T, OHSAKI Y, SUZUKI M, et al.Inhibition of Niemann-Pick-type C1-like1 by ezetimibe activates autophagy in human hepatocytes and reduces mutant α1-antitrypsin Z deposition[J]. Hepatology, 2014, 59(4): 1591-1599.
[5] MORTIMORE G E, HUTSON N J, SURMACZ C A. Quantitative correlation between proteolysis and macro-andmicroautophagy in mouse hepatocytes during starvation and refeeding[J]. Proceedings of the National Academy of Sciences, 1983, 80(8): 2179-2183.
[6] TECKMAN J H, MANGALAT N. Alpha-1 antitrypsin and liver disease: mechanisms of injury and novel interventions[J]. Expert Review of Gastroenterology and Hepatology, 2015, 9(2): 261-268.
[7] KOMANDER D, RAPE M. The ubiquitin code[J]. Annual Review of Biochemistry, 2012, 81(7): 203-229.
[8] HARRIS S L, LEVINE A J. The p53 pathway: positive and negative feedback loops[J]. Oncogene, 2005, 24(17): 2899-2908.
[9] SOWA M E, BENNETT E J, GYGI S P, et al. Defining the human deubiquitinating enzyme interaction landscape[J]. Cell, 2009, 138(2): 389-403.
[10] KESSLER B M, FORTUNATI E, MELIS M, et al. Proteome changes induced by knock-down of the deubiquitylating enzyme HAUSP/USP7[J]. Journal of Proteome Research, 2007, 6(11): 4163-4172.
[11] KON N, KOBAYASHI Y, LI M, et al. Inactivation of HAUSP in vivo modulates p53 function[J]. Oncogene, 2010, 29(9): 1270-1279.
[12] SOWA M E, BENNETT E J, GYGI S P, et al. Defining the human deubiquitinating enzyme interaction landscape[J]. Cell, 2009, 138(2): 389-403.
[13] 高冰芳, 杨悦, 杨京京, 等. Dnmt1, Caveolin-1在结直肠癌中的表达及其临床意义[J]. 辽宁医学院学报, 2011, 32(1): 9-12.
[14] SACCO J J, COULSON J M, CLAGUE M J, et al. Emerging roles of deubiquitinases in cancer-associated pathways[J]. IUBMB Life, 2010, 62(2): 140-157.
[15] MATTHEWS J R, NICHOLSON J, JAFFRAY E, et al. Conformational changes induced by DNA binding of NF-κB[J]. Nucleic Acids Research, 1995, 23(17): 3393-3402.
[16] COLLERAN A, COLLINS P E, O’ CARROLL C, et al. Deubiquitination of NF-κB by ubiquitin-specific protease-7 promotes transcription[J]. Proceedings of the National Academy of Sciences, 2013, 110(2): 618-623.
[17] NAGAI M, IKEDA S. Carbohydrate metabolism in fish:Ⅰ. Effects of starvation and dietary composition on the blood glucose level and the hepatopancreatic glycogen and lipid contents in carp[J]. Bulletin of the Japanese Society of Scientific Fisheries, 1971, 37(5): 404-409.
[18] INCE B W, THORPE A.The in vitro metabolism of 14C-glucose and 14C-glycine in insulin-treated Northern pike (Esox lucius L.) [J]. General and Comparative Endocrinology,1976, 28(4): 481-486.
[19] FORAND A, KOUMAKIS E, ROUSSEAU A, et al. Disruption of phosphate transporter Pit1 in hepatocytes improves glucose metabolism and insulin signaling by modulating the USP7/IRS1 interaction[J]. Cell Reports, 2016, 16(10):2736-2748.
[20] JIANG L, XIONG J, ZHAN J, et al. Ubiquitin-specific peptidase 7 (USP7)-mediated deubiquitination of the histone deacetylase SIRT7 regulates gluconeogenesis[J]. Journal of Biological Chemistry, 2017, 292(32):13296-13311.
[21] HALL J A, TABATA M, RODGERS J T, et al. USP7 attenuates hepatic gluconeogenesis through modulation of FoxO1 gene promoter occupancy[J]. Molecular Endocrinology, 2014, 28(6): 912-924.
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