旋毛虫plancitoxin-1-like基因的序列分析与编码蛋白结构预测

doi:10.11843/j.issn.0366-6964.2018.01.025

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (1): 218-222.doi: 10.11843/j.issn.0366-6964.2018.01.025

• 研究简报 • 上一篇

旋毛虫plancitoxin-1-like基因的序列分析与编码蛋白结构预测

廖成水^1,4, 王晓利², 田文静¹, 杨亚东¹, 张梦珂¹, 王安琪⁴, 张春杰¹, 刘明远⁴, Boireau Pascal^4,5, 程相朝^1,3*

1. 河南科技大学, 动物科技学院/洛阳市活载体生物材料与动物疫病防控重点实验室, 洛阳 471023;
2. 河南科技大学医学院, 洛阳 471023;
3. 洛阳职业技术学院, 洛阳 471003;
4. 吉林大学人兽共患病研究所/人兽共患病研究教育部重点实验室, 长春 130062;
5. 法国国家食品、环境及劳动卫生署动物卫生实验室, 迈松阿尔福 94706

收稿日期:2017-06-26 出版日期:2018-01-23 发布日期:2018-01-18
通讯作者: 程相朝,教授,主要从事动物疫病分子与免疫学机制研究,E-mail:chengxch@126.com
作者简介:廖成水(1986-),男,博士,主要从事微生物感染与先天性免疫细胞作用研究,E-mail:liaochengshui33@163.com
基金资助:
国家自然科学基金（31030064）；国家自然科学基金面上项目（31572489）；河南科技大学博士启动基金（13480071）；河南省高等学校重点科研项目计划（17A230009）

Cloning and Sequence Analysis of Plancitoxin-1-like Gene in Trichinella spiralis

LIAO Cheng-shui^1,4, WANG Xiao-li², TIAN Wen-jing¹, YANG Ya-dong¹, ZHANG Meng-ke¹, WANG An-qi⁴, ZHANG Chun-jie¹, LIU Ming-yuan⁴, BOIREAU Pascal^4,5, CHENG Xiang-chao^1,3*

1. College of Animal Science and Technology/Luoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang 471023, China;
2. Medical College, Henan University of Science and Technology, Luoyang 471023, China;
3. Luoyang Vocational and Technical College, Luoyang 471003, China;
4. Key Laboratory of Zoonisis of Ministry of Education, Institute of Zoonosis, Jilin University, Changchun 130062, China;
5. ANSES's Laboratory for Animal Health, Maisons-Alfort 94706, France

Received:2017-06-26 Online:2018-01-23 Published:2018-01-18

摘要/Abstract

摘要：

基因组预测旋毛虫拥有125种庞大的DNase Ⅱ家族蛋白，但只有plancitoxin-1唯一在C端含有一个保守的DNase Ⅱ活性位点（HKD基序）。为研究旋毛虫plancitoxin-1基因的序列结构特征，以旋毛虫总RNA为模板进行RT-PCR，获得plancitoxin-1基因的序列，并利用生物信息学方法进行系统性分析。结果显示，扩增得到的核苷酸序列比GenBank中相关序列短210 bp，编码294个氨基酸，由20种氨基酸组成，重新命名为plancitoxin-1-like，且氨基酸序列中在N端和C端都具有HKD基序。蛋白质理论相对分子质量为33.19 ku，等电点为9.17，是稳定存在的亲水性蛋白。Plancitoxin-1-like的氨基酸序列与其他物种DNase Ⅱ的相似性为35%左右。Plancitoxin-1-like含有4个N-糖基化位点、3个O-糖基化位点、31个磷酸化位点、17个B细胞线性结合位点和9个T细胞结合位点以及3个二硫键。二级结构中，α-螺旋占14.29%（42个），延伸链占28.23%（83个），无规卷曲占55.10%（162个）。本研究成功克隆旋毛虫plancitoxin-1-like基因，对其序列结构进行分析，为进一步研究庞大的DNase Ⅱ家族蛋白在旋毛虫发育和感染中的作用奠定基础。

关键词: 旋毛虫, DNase II, 克隆, 生物信息学

Abstract:

It is noteworthy that more extensive expansion of the DNase Ⅱ-like protein family (estimated 125 genes) in the genome of the parasitic nematode Trichinella spiralis (T. spiralis). However, only plancitoxin-1 possesses one predicted active site of DNase Ⅱ (HKD motif) in its C-terminus domain in 125 DNase Ⅱ-like protein family. To explore the molecular characteristics of plancitoxin-1 gene in T. spiralis, plancitoxin-1 gene was cloned by RT-PCR from total RNA of T. spiralis larvae, and analyzed with bioinformatics software. The results showed that the sequences of plancitoxin-1 cloned from T. spiralis were shorter (210 bp less) than the XM_003370715.1 sequence in GenBank, and named after plancitoxin-1-like. The gene encoded a protein of 294 amino acids. Intriguingly, there were two HKD motifs in the N-and C-termini in the cloned sequences. plancitoxin-1-like is a stable protein with a predicted molecular mass (Mr) of 33.19 kDa and theoretical pI of 9.17. By sequence alignment, plancitoxin-1-like protein revealed 35% sequence identity to previously reported DNase Ⅱ family members from other species. A total of 4 N-linked glycosylation sites, 3 O-linked glycosylation sites, 31 phosphorylation site 17 linear B cell piptope, 9 T cell binding sites, and 3 disulfide bond were predicted in plancitoxin-1-like. Analysis of secondary structure revealed that α-helix, extended strand, and random coil were 14.29% (42), 28.23% (83), and 55.10% (162), respectively. In this study, the plancitoxin-1-like gene was cloned and analyzed. The results laid the foundation for the further study on the role of development infection in T. spiralis.

Key words: Trichinella spiralis, DNase Ⅱ, clone, bioinformatics

中图分类号:

S852.731

廖成水, 王晓利, 田文静, 杨亚东, 张梦珂, 王安琪, 张春杰, 刘明远, Boireau Pascal, 程相朝. 旋毛虫plancitoxin-1-like基因的序列分析与编码蛋白结构预测[J]. 畜牧兽医学报, 2018, 49(1): 218-222.

LIAO Cheng-shui, WANG Xiao-li, TIAN Wen-jing, YANG Ya-dong, ZHANG Meng-ke, WANG An-qi, ZHANG Chun-jie, LIU Ming-yuan, BOIREAU Pascal, CHENG Xiang-chao. Cloning and Sequence Analysis of Plancitoxin-1-like Gene in Trichinella spiralis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 218-222.

参考文献

[1] CUNNINGHAM L, LASKOWSKI M. Presence of two different desoxyribonucleode-polymerases in veal kidney[J]. Biochim Biophys Acta, 1953, 11(4):590-591.
[2] YASUDA T, TAKESHITA H, ⅡDA R, et al. Molecular cloning of the cDNA encoding human deoxyribonuclease Ⅱ[J]. J Biol Chem, 1998, 273(5):2610-2616.
[3] HOU N, PIAO X, CAI P, et al. A novel Schistosoma japonicum endonuclease homologous to DNase Ⅱ[J]. BMC Genomics, 2015, 16:126.
[4] SHIMONI Z, FROOM P. Uncertainties in diagnosis, treatment and prevention of trichinellosis[J]. Expert Rev Anti Infect Ther, 2015, 13(10):1279-1288.
[5] MITREVA M, JASMER D P, ZARLENGA D S, et al. The draft genome of the parasitic nematode Trichinella spiralis[J]. Nat Genet, 2011, 43(3):228-235.
[6] YU H, LAI H J, LIN T W, et al. Autonomous and non-autonomous roles of DNase Ⅱ during cell death in C. elegans embryos[J].Biosci Rep,2015,35(3).pii:e00203.
[7] SCHÄFER P, CYMERMAN I A, BUJNICKI J M, et al. Human lysosomal DNase Ⅱalpha contains two requisite PLD-signature (HxK) motifs:evidence for a pseudodimeric structure of the active enzyme species[J]. Protein Sci, 2007, 16(1):82-91.
[8] MAK C H, KO R C. Characterization of endonuclease activity from excretory/secretory products of a parasitic nematode, Trichinella spiralis[J]. Eur J Biochem, 1999, 260(2):477-481.
[9] BIEN J, NÄREAHO A, VARMANEN P, et al. Comparative analysis of excretory-secretory antigens of Trichinella spiralis and Trichinella britovi muscle larvae by two-dimensional difference gel electrophoresis and immunoblotting[J]. Proteome Sci, 2012, 10(1):10.
[10] VARELA-RAMIREZ A, ABENDROTH J, MEJIA A A, et al. Structure of acid deoxyribonuclease[J]. Nucleic Acids Res, 2017, 45(10):6217-6227.
[11] JASMER D P, KWAK D. Fusion and differentiation of murine C2C12 skeletal muscle cells that express Trichinella spiralis p43 protein[J]. Exp Parasitol, 2006, 112(2):67-75.
[12] VASSILATIS D K, DESPOMMIER D, MISEK D E, et al. Analysis of a 43-kDa glycoprotein from the intracellular parasitic nematode Trichinella spiralis[J]. J Biol Chem, 1992, 267(26):18459-18465.
[13] MACLEA K S, KRIESER R J, EASTMAN A. A family history of deoxyribonuclease Ⅱ:surprises from Trichinella spiralis and Burkholderia pseudomallei[J]. Gene, 2003, 305(1):1-12.

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旋毛虫plancitoxin-1-like基因的序列分析与编码蛋白结构预测

Cloning and Sequence Analysis of Plancitoxin-1-like Gene in Trichinella spiralis

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