黄鳍鲷肌肉中弹性蛋白酶基因全长克隆及生物信息学
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

Q785;S965.3

基金项目:

国家自然科学基金(31701676,31760483);江西省科技厅自然科学基金(20192BAB214026);江西省教育厅科技计划重点项目(GJJ180173)


Cloning and bioinformatic analysis of elastase gene from skeletal muscle of yellowfin seabream (Acanthopagrus latus)
Author:
Affiliation:

Fund Project:

National Natural Science Foundation of China (31701676, 31760483); Natural Science Foundation of Jiangxi Province (20192BAB214026); .Jiangxi Province Department of Education Science and technology research project (GJJ180173)

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    研究表明弹性蛋白酶(elastase, Ela)参与调控嗜水气单胞菌引起的细菌性溃疡病,对控制鱼类养殖中免疫疾病有重要作用。本研究根据已报道脊椎动物的Elas基因序列设计特异性引物,通过RT-PCR和RACE技术克隆获得黄鳍鲷Ela基因(AlEla)cDNA序列全长,并对其进行生物信息学分析。结果显示,AlEla基因全长为946 bp,包含807 bp的开放阅读框,编码269个氨基酸,含有16个氨基酸信号肽的亲水性稳定膜外蛋白,定位于细胞外基质,具有3个O-糖基化和15个磷酸化的潜在位点。编码的成熟AlEla蛋白预测相对分子质量27 509 u,等电点为6.05。氨基酸序列同源性比对发现,AlEla与鲷和鲆的Elas2的同源性为84.76%~99.26%,处于系统进化树上同一分支,表明它们的亲缘性相近。AlEla具有Elas的保守结构,如Tryp-SPc结构域、丝氨酸蛋白酶催化三联体、保守的半胱氨酸残基以及在C端含有GDSGGPL序列等,揭示该酶有丝氨酸蛋白酶的催化活性。对AlEla蛋白的二级结构分析显示,α-螺旋、β-转角、β-折叠和无规则卷曲所占的比例分别为17.84%、7.06%、24.54%和50.56%。三级结构分析推测α-螺旋和β-折叠对于AlEla蛋白的空间构象可能有着重要作用。本研究从黄鳍鲷肌肉中成功克隆AlEla基因并分析其生物信息学特征,为后续研究AlEla参与调控鱼类免疫疾病的机制提供理论基础。

    Abstract:

    Researches show that elastase (Ela) is involved in regulation of bacterial ulcer caused by Aerononas hydrophila, and plays an important role in control of immune diseases during fish culture. The full-length cDNA sequence of elastase gene was cloned from Acanthopagrus latus based on the reported sequencing data of other vertebrates by using RT-PCR and RACE, which was further analyzed by bioinformatics analysis. The cloned AlEla gene was 946 bp, containing a 807 bp open reading frame which encoded 269 amino acids. AlEla protein was a hydrophilic stable extracellular protein containing a signal peptide with 16 amino acid residues, which might be located in extracellular matrix. Furthermore, AlEla protein has 3 predicted O-glycosylation sites and 15 predicted phosphorylation sites. The theoretical molecular weight of mature protein was 27 509 u and its isoelectric point was 6.05. The analysis of amino acid sequence homology showed that it has high similarity with Sparus aurata, Spondyliosoma cantharus, Pagrus major, Scophthalmus maximus, Paralichthys olivaceus (84.76%-99.26%). They were also in the same branch in phylogenetic tree, indicating that they are the close relatives. Moreover, AlEla has the conserved structures of Elas, including conserved region Tryp-SPc superfamily, catalytic triad of serine proteinase active site, conserved cysteine residues and the sequence of “GDSGGPL” in C-terminal, which revealed that AlEla might show serine proteinase catalytic activity. In the secondary structure of AlEla protein, the proportion of alpha helix, beta turn, extended strand and random coil was 17.84%, 7.06%, 24.54% and 50.56%, respectively. The result of predicted tertiary structure of AlEla indicated that alpha helix and extended strand might play an important role in its conformation structure. In the present study, AlEla gene was successfully cloned from Acanthopagrus latus and we analyzed its bioinformatics, providing theoretical basis for further elucidation of mechanism of regulation of fish immune diseases by AlEla protein.

    参考文献
    相似文献
    引证文献
引用本文

钟婵,吴国平,舒梅,孙乐常,曹敏杰.黄鳍鲷肌肉中弹性蛋白酶基因全长克隆及生物信息学[J].水产学报,2021,45(1):125~135

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2019-11-23
  • 最后修改日期:2020-06-23
  • 录用日期:2020-07-02
  • 在线发布日期: 2021-01-08
  • 出版日期: