半滑舌鳎细胞分裂周期蛋白42基因克隆与结构预测

doi:10.19640/j.cnki.jtau.2022.03.007

天津农学院学报 ›› 2022, Vol. 29 ›› Issue (3): 36-42.doi: 10.19640/j.cnki.jtau.2022.03.007

半滑舌鳎细胞分裂周期蛋白42基因克隆与结构预测

刘厚孚¹, 谭静¹, 胡秀彩¹, 管振国², 吕爱军^{1,通信作者}, 孙敬锋¹

1.天津农学院水产学院天津市水产生态及养殖重点实验室,天津 300392;
2.天津鼎正新兴生物技术有限公司,天津 300383

收稿日期:2020-07-05 出版日期:2022-09-30 发布日期:2022-10-10
通讯作者: 吕爱军（1973—）,男,教授,博士,主要从事水产动物微生物与免疫学研究。E-mail：lajand@126.com。
作者简介:刘厚孚（1998—）,男,硕士在读,主要从事水产动物微生物与免疫学研究。E-mail：1055688654@qq.com。
基金资助:
天津市自然科学基金项目（18JCYBJC29900）; 天津市技术创新引导专项基金（21YDPJC00510）; 天津市高校重点科研项目（2019ZD14）

Cloning and structure prediction of cell division cyclin 42 gene from Cynoglossus semilaevis

Liu Houfu¹, Tan Jing¹, Hu Xiucai¹, Guan Zhenguo², Lü Aijun^{1,Corresponding Author}, Sun Jingfeng¹

1. Tianjin Key Laboratory of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300392, China;
2. S&E Burgeoning Biotechnology（Tianjin）Co., Ltd., Tianjin 300383, China

Received:2020-07-05 Online:2022-09-30 Published:2022-10-10

摘要/Abstract

摘要： 细胞分裂周期蛋白42（Cdc42）是Rho超家族酶中的一个亚类,具有调节细胞骨架和肌动蛋白作用,并参与免疫与炎症反应。设计半滑舌鳎Cdc42基因（即CsCdc42）特异性引物,采用RT-PCR方法、生物信息学方法等进行CsCdc42基因克隆及结构预测。结果获得CsCdc42基因片段长度为847 bp,推测编码191个氨基酸,含有7个抗原表位（27-28、61-63、90-92、120-122、131-133、164-166和181-183 aa）、1个RHO结构域（6-179 aa）,其中含有1个ATP/GTP结合位点和1个酰基化位点;蛋白结构预测主要为α螺旋（30.9%）、β折叠（24.6%）和无规则卷曲（44.5%）。序列比对结果表明,CsCdc42与斑马鱼、青鳉、虹鳟、大菱鲆和虾等氨基酸序列同源性分别为99.43%、97.91%、95.81%、94.76%和91.10%;构建系统发育树显示CsCdc42与斑马鱼、人和海水鱼类Cdc42亲缘关系最近,自然聚为一支。以上结果为研究半滑舌鳎细胞分裂周期蛋白相关基因功能提供科学参考。

关键词: 半滑舌鳎, 细胞分裂周期蛋白42, 基因克隆, 结构预测

Abstract: Cdc42 is a subclass of Rho superfamily enzymes that regulates cytoskeleton and actin activity, and participates in immune and inflammatory responses. In this study, the specific primers were designed for Cdc42 gene（CsCdc42）of Cynogl ossus semilaevis, and its cloning and structure prediction were performed by RT-PCR and bioinformatics methods. The results showed that the fragment of CsCdc42 gene was 847 bp in length, encoding 191 amino acids（aa）, and contained 7 epitopes （27-28, 61-63, 90-92, 120-122, 131-133, 164-166 and 181-183 aa）, one RHO domain（6-179 aa）, ATP/GTP binding site and acylation site, respectively. The protein structure was predicted by helix（30.9%）, β-fold（24.6%）and irregular coil（44.5%）. The results of sequence alignment showed that the homology of amino acid sequences of CsCdc42 was 99.43%, 97.91%, 95.81%, 94.76% and 91.10%, respectively, for zebrafish（Danio rerio）, medaka（Oryzias latipes）, rainbow trout（Oncorhynchus mykiss）, turbot （Scophthalmus maximus）and shrimp（Penaeus vannamei）. Phylogenetic tree analysis showed that Cdc42 was closely related to zebrafish（Danio rerio）, human（Homo sapiens）and several marine fishes, and naturally grouped into one cluster. These results provide a scientific reference for further study of the function of cyclin-related genes in Cynoglossus semilaevis.

Key words: Cynoglossus semilaevis, Cdc42, gene cloning, structure prediction

中图分类号:

S917.4

刘厚孚, 谭静, 胡秀彩, 管振国, 吕爱军, 孙敬锋. 半滑舌鳎细胞分裂周期蛋白42基因克隆与结构预测[J]. 天津农学院学报, 2022, 29(3): 36-42.

Liu Houfu, Tan Jing, Hu Xiucai, Guan Zhenguo, Lü Aijun, Sun Jingfeng. Cloning and structure prediction of cell division cyclin 42 gene from Cynoglossus semilaevis[J]. Journal of Tianjin Agricultural University, 2022, 29(3): 36-42.

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: http://xuebao.tjau.edu.cn/CN/10.19640/j.cnki.jtau.2022.03.007

http://xuebao.tjau.edu.cn/CN/Y2022/V29/I3/36

参考文献

[1] HAN Z R,SUN J F,LU A J,et al.Identification and characterization of Shewanella algae from reared tongue sole,Cynoglossus semilaevis Günther[J]. Aquaculture,2017,468(2017):356-362.
[2] 邓思平,陈松林,田永胜,等. 半滑舌鳎的性腺分化和温度对性别决定的影响[J]. 中国水产科学,2007,14(5):714-719.
[3] 廉正义,胡秀彩,吕爱军,等. 半滑舌鳎黏蛋白MUC18基因克隆与序列分析[J]. 水产学杂志,2019(4):23-30.
[4] MARIA C F,KARIN D,ANJA N,et al.Cdc42 activity regulates hematopoietic stem cell aging and rejuvenation[J]. Cell Stem Cell,2012,10(5):520-530.
[5] 李丽丽,赵超,范嗣刚,等. 斑节对虾CdC42基因的克隆及其在不同胁迫条件下的表达分析[J]. 南方水产科学,2018,14(4):37-45.
[6] KOH C G,LIM L.Cdc42 and rac control of the actin cytoskeleton[J]. Rho Family GTPases,2005,3:137-156.
[7] DOUGLAS I J.Cdc42:An essential Rho-type GTPase conorolling eukaryotic cell polarity[J]. microbiol Mol Biol Rev,1999,63(1):54-105.
[8] KAIBUCHI K,KURODA S,AMANO M.Regulation of the cytoskeleton and cell adhesion by the Rho family Gtpases in mammalian cells[J]. Annual Review of Biochemistry,1999,68(1):459-486.
[9] LU X,WU X,ANA P,et al.Cdc42 and Rac1 are implicated in the activation of the Nef-associated kinase and replication of HIV-1[J]. Current Biology,1996,6(12):1677-1684.
[10] BURBAGE M,KEPPLER S J,GASPARRINI F,et al.Cdc42 is a key regulator of B cell differentiation and is required for antiviral humoral immunity[J]. Journal of Experimental Medicine,2015,212(1):53-72.
[11] HADDAD E,ZUGAZA J L,LOUACHE F,et al.The interaction between Cdc42 and WASP is required for SDF-1- induced T-lymphocyte chemotaxis[J]. Blood,2001,97(1):33-38.
[12] WESTERBERG L,GREICIUS G,SNAPPER S B,et al.Cdc42,Rac1,and the Wiskott-Aldrich syndrome protein are involved in the cytoskeletal regulation of B lymphocytes[J]. Blood,2001,98(4):1086-1094.
[13] 王媛. 巨噬细胞特异性敲除Cdc42基因加重细菌脂多糖LPS诱导的小鼠炎症反应[D]. 南昌:南昌大学,2016.
[14] 胡默俨. 草鱼两个GTPase基因的分子克隆及免疫功能分析[D]. 上海:上海海洋大学,2016.
[15] HUSAIN M,BIRD S,ZWIETEN R V,et al.Cloning of the IL-1β3 gene and IL-1β4 pseudogene in salmonids uncovers a second type of IL-1β gene in teleost fish[J]. Developmental & Comparative Immunology,2012,38(3):431-446.
[16] 舒欢欢,郑伟贤,钱云霞. 鲈鱼ZnT基因的cDNA克隆及生物信息学分析[J]. 生物学杂志,2015(1):6-9.
[17] 谭静,胡秀彩,吕爱军,等. 半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测[J]. 天津农学院学报,2019,26(2):51-56.
[18] LI J,LIU Y,JIN Y,et al.Essential role of Cdc42 in cardiomyocyte proliferation and cell-cell adhesion during heart development[J]. Developmental Biology,2016,421(2):271.
[19] OLENIK C,BARTH H,JUST I,et al.Gene expression of the small GTP-binding proteins RhoA,RhoB,Rac1,and Cdc42 in adult rat brain[J]. Molecular Brain Research,1997,52(2):269.
[20] ZAN L,HUI L I,WEN-JING M A,et al. Expression and Significance of Cdc42 and Rb Gene in Kazak’s Esophageal Cancer[J]. Endemic Diseases Bulletin,2008,4:1-3.
[21] CHERNICHENKO N,OMELCHENKO T,DEBORDE S,et al.Abstract 460:The role of Cdc42 in cancer cell perineural invasion[J]. Cancer Research,2012,72(8):460.

半滑舌鳎细胞分裂周期蛋白42基因克隆与结构预测

Cloning and structure prediction of cell division cyclin 42 gene from Cynoglossus semilaevis

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 11

编辑推荐

Metrics

本文评价

[1]	杨艳凤, 王庆奎, 石洪玥, 徐大为, 孙学亮. 养殖与野生半滑舌鳎肌肉营养成分对比分析[J]. 天津农学院学报, 2024, 31(2): 56-62.
[2]	马国宁, 田云臣, 胡金城, 赵晴晴, 隋金柱, 郑杰. 一家典型海水工厂化循环水养殖系统运行效果分析——以半滑舌鳎为例[J]. 天津农学院学报, 2024, 31(2): 71-77.
[3]	聂国梁, 王庆奎, 赵鸿昊, 王祯辉, 付建东, 李强. 溶解氧对半滑舌鳎呼吸频率、生理生化指标和组织结构的影响[J]. 天津农学院学报, 2024, 31(1): 54-60.
[4]	王妍, 王庆奎, 方珍珍, 石洪玥, 吴京霞, 郑海玲. 半滑舌鳎幼鱼不同阶段肠道和肝脏的组织切片观察[J]. 天津农学院学报, 2023, 30(6): 32-40.
[5]	葛仑华, 江博赟, 孙敬锋, 吕爱军, 胡秀彩, 陈丽梅. 三种DNA提取方法对半滑舌鳎肠道菌群高通量测序结果的影响[J]. 天津农学院学报, 2022, 29(3): 29-35.
[6]	罗希帆, 周灵美, 谢晓东, 陈帅君, 杨红澎, 吴疆. 一氧化氮合成酶的基因克隆与功能鉴定[J]. 天津农学院学报, 2022, 29(2): 13-17.
[7]	谭静, 胡秀彩, 吕爱军, 孙敬锋, 刘小雪, 廉正义. 半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测[J]. 天津农学院学报, 2019, 26(2): 51-56.
[8]	葛婧, 郭永军, 王庆奎, 徐大为, 孙学亮, 陈成勋, 邢克智. 连翘、黄连、穿心莲对半滑舌鳎血液抗氧化指标的影响[J]. 天津农学院学报, 2016, 23(4): 21-27.
[9]	方珍珍, 王庆奎, 陈成勋, 郭永军, 邢克智. 复方中草药对半滑舌鳎和盾纤毛虫的急性毒性研究[J]. 天津农学院学报, 2015, 22(4): 6-8.
[10]	胡璇, 孙敬锋, 陈成勋, 邢克智. 养殖半滑舌鳎腹水病的病原分离鉴定及药物敏感性分析[J]. 天津农学院学报, 2014, 21(3): 12-16.
[11]	赵从明, 刘刚. 养殖密度对半滑舌鳎成鱼生长的影响[J]. 天津农学院学报, 2013, 20(3): 34-36.