斯氏副柔线虫CAMK/TSSK蛋白激酶基因的克隆及其蛋白质结构与抗原表位预测分析

doi:10.11843/j.issn.0366-6964.2018.03.023

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (3): 644-651.doi: 10.11843/j.issn.0366-6964.2018.03.023

斯氏副柔线虫CAMK/TSSK蛋白激酶基因的克隆及其蛋白质结构与抗原表位预测分析

王文龙¹, 赵学亮¹, 孙柯¹, 冯陈晨¹, 白丽艳¹, 王姝懿^1,3, 王梦雅², 刘春霞^4*

1. 内蒙古农业大学兽医学院, 农业部动物疾病临床诊疗技术重点实验室, 呼和浩特 010018;
2. 内蒙古农业大学动物科学学院, 呼和浩特 010018;
3. 内蒙古自治区综合疾病预防控制中心, 呼和浩特 010018;
4. 内蒙古农业大学生命科学学院, 呼和浩特 010018

收稿日期:2017-10-19 出版日期:2018-03-23 发布日期:2018-03-13
通讯作者: 刘春霞,博士,副教授,E-mail:lcx.imau@163.com
作者简介:王文龙(1976-),内蒙古人,博士,副教授,主要从事寄生虫组学与分子免疫学研究,E-mail:wwl.imau@163.com;赵学亮(1992-),男,河北沧州人,硕士生,主要从事寄生虫组学及分子免疫学研究,E-mail:zhaoxueliang92@sina.com。二人并列第一作者
基金资助:
国家自然科学基金（31260603）；内蒙古自然科学基金（2016MS0341）

Cloning, Protein Structure Analysis and Antigenic Epitopes Prediction of Immune-related Gene, CAMK/TSSK Protein Kinase Gene of Parabronema skrjabini

WANG Wen-long¹, ZHAO Xue-liang¹, SUN Ke¹, FENG Chen-chen¹, BAI Li-yan¹, WANG Shu-yi^1,3, WANG Meng-ya², LIU Chun-xia^4*

1. Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease of Ministry of Agriculture, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China;
3. Inner Mongolia Center for Disease Control and Prevention, Hohhot 010018, China;
4. College of Life Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received:2017-10-19 Online:2018-03-23 Published:2018-03-13

摘要/Abstract

摘要：

为分析并预测斯氏副柔线虫免疫相关基因CTPK编码蛋白结构、功能以及其作为疫苗候选抗原的可能性，通过提取斯氏副柔线虫组织RNA，反转录合成cDNA，设计特异性引物以扩增CTPK基因的CDS区，并利用生物信息学软件对CTPK进行蛋白质结构分析和抗原表位预测。生物信息学分析表明，CTPK基因cDNA序列全长共519 bp，具有完整的开放阅读框（519 bp），编码173个氨基酸，相对分子质量和等电点大小分别为20.264 ku和9.53。结构分析发现，蛋白质无跨膜区，无规则卷曲构成二级结构的主要组分，亲水性氨基酸比例超过60%；抗原表位预测表明，CTPK蛋白是一种抗原性较高的亲水性蛋白，既含有较多潜在的B细胞抗原表位又存在较多的T细胞抗原表位。推测CTPK有望用作斯氏副柔线虫的免疫诊断抗原和疫苗候选抗原，本研究为骆驼斯氏副柔线虫生前诊断方法iELISA的建立和DNA疫苗的研究奠定了理论基础。

关键词: 斯氏副柔线虫, CTPK基因, 抗原表位, 蛋白质结构预测, 生物信息学分析, 丝/苏氨酸蛋白激酶

Abstract:

Our study was conducted to explore protein structure, function and the possibility as a vaccine candidate antigen of immune-related gene, CAMK/TSSK protein kinase (CTPK) gene of Parabronema skrjabini. RNA was extracted from Parabronema skrjabini and cDNA was obtained by reverse transcription. A pair of specific primers was designed to amplify the coding region of CTPK gene. The protein structure and antigenic epitopes of CTPK were analyzed by using bioinformatics softwares. The bioinformatic analysis results showed that the length of CTPK gene was 519 bp, contained an open reading frame of 519 bp which encoded a polypeptide of 173 amino acids with a predicted molecular mass of 20.264 kDa and PI of 9.53. Analyses of domain and structure indicated that the protein was dominantly composed of 60% hydrophillic amino acid residues, with numerous curls and no transmembrane structure. The antigenic epitopes prediction results indicated that CTPK protein is a hydrophilic protein with high antigenicity. It contained more potential B cell antigen epitopes and more T cell antigen epitopes. These results possibly suggested that the CTPK antigen has been promised as a candidate for immune diagnostics and could be used as a vaccine against Parabronema skrjabini. This research provide the theoretical basis in order to establish iELISA corresponding diagnostic kits and development DNA vaccine.

Key words: Parabronema skrjabini, CTPK gene, antigenic epitopes, protein structure prediction, bioinformation analysis, serine/threonine protein kinase

中图分类号:

S852.731

王文龙, 赵学亮, 孙柯, 冯陈晨, 白丽艳, 王姝懿, 王梦雅, 刘春霞. 斯氏副柔线虫CAMK/TSSK蛋白激酶基因的克隆及其蛋白质结构与抗原表位预测分析[J]. 畜牧兽医学报, 2018, 49(3): 644-651.

WANG Wen-long, ZHAO Xue-liang, SUN Ke, FENG Chen-chen, BAI Li-yan, WANG Shu-yi, WANG Meng-ya, LIU Chun-xia. Cloning, Protein Structure Analysis and Antigenic Epitopes Prediction of Immune-related Gene, CAMK/TSSK Protein Kinase Gene of Parabronema skrjabini[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 644-651.

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斯氏副柔线虫CAMK/TSSK蛋白激酶基因的克隆及其蛋白质结构与抗原表位预测分析

Cloning, Protein Structure Analysis and Antigenic Epitopes Prediction of Immune-related Gene, CAMK/TSSK Protein Kinase Gene of Parabronema skrjabini

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