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

Abstract

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.

CLC Number:

S852.731

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.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2018.03.023

https://www.xmsyxb.com/EN/Y2018/V49/I3/644

References

[1] 卢俊杰, 靳家声. 人和动物寄生线虫图谱[M]. 北京:中国农业科学技术出版社, 2002:405-459.
LU J J, JIN J S. The map of human and animal parasitic nematodes[M]. Beijing:China Agricultural Science and Technology Press, 2002:405-459. (in Chinese)
[2] 沈杰, 黄兵. 中国家畜家禽寄生虫名录[M]. 北京:中国农业科学技术出版社, 2004:105-111.
SHEN J, HUANG B. A list of parasites for livestock and poultry in China[M]. Beijing:China Agricultural Science and Technology Press, 2004:105-111. (in Chinese)
[3] HASHEMINASAB S S. Molecular characterization of the first internal transcribed spacer of rDNA of Parabronema skrjabini for the first time in sheep[J]. Ann Parasitol, 2015, 61(4):241-246.
[4] HASHEMINASAB S S, JALOUSIAN F, MESHGI B. Molecular and morphological characterization of Parabronema skrjabini of sheep and goats at three different geographical zones in Iran[J]. Ann Parasitol, 2016, 62(1):55-61.
[5] 杨晓野. 内蒙古自治区骆驼寄生虫种类调查[D]. 呼和浩特:内蒙古农牧学院, 1985:40-41.
YANG X Y. Camel parasitic species investigation in Inner Mongolia Autonomous Region[D]. Hohhot:Inner Mongolia Agricultural University, 1985:40-41. (in Chinese)
[6] 杨莲茹, 杨晓野, 刘珍莲, 等. 内蒙古地区骆驼斯氏副柔线虫(Parabronema skrjabini)病调查[J]. 内蒙古农业大学学报, 2004, 25(1):43-45.
YANG L R, YANG X Y, LIU Z L, et al. Investigation of Parabronema Skrjabini disease of camels in Inner Mongolia Region[J]. Journal of Inner Mongolia Agricultural University, 2004, 25(1):43-45. (in Chinese)
[7] 赵治国. 我国骆驼斯氏副柔线虫病传播媒介的研究[D]. 呼和浩特:内蒙古农业大学, 2010.
ZHAO Z G. Study on the vector of camel Parabronemosis in China[D]. Hohhot:Inner Mongolia Agricultural University, 2010. (in Chinese)
[8] 张晓东, 杨晓野, 杨莲茹, 等. 斯氏副柔线虫rDNA-ITS片段的克隆及序列分析[J]. 畜牧兽医学报, 2009, 40(5):775-779.
ZHANG X D, YANG X Y, YANG L R, et al. Cloning and sequence analysis of the rDNA-ITS of Parabronema skrjabini[J]. Acta Veterinaria et Zootechnica Sinica, 2009, 40(5):775-779. (in Chinese)
[9] 冯陈晨, 王文龙, 呼和巴特尔. 骆驼斯氏副柔线虫转录组的高通量测序及分析[J]. 中国兽医学报, 2017, 37(4):671-675.
FENG C C, WANG W L, HUHR B T E. Sequencing and analysis of the transcriptome of Parabronema skrjabini in camle[J]. Chinese Journal of Veterinary Science, 2017, 37(4):671-675. (in Chinese)
[10] SIMÕES A E S, PEREIRA D M, AMARAL J D, et al. Efficient recovery of proteins from multiple source samples after trizol^® or trizol^®·LS RNA extraction and long-term storage[J]. BMC Genomics, 2013, 14:181.
[11] PETERSEN T N, BRUNAK S, VON HEIJNE G, et al. SignalP 4.0:discriminating signal peptides from transmembrane regions[J]. Nat Methods, 2011, 8(10):785-786.
[12] KELLEY L A, MEZULIS S, YATES C M, et al. The Phyre2 web portal for protein modeling, prediction and analysis[J]. Nat Protoc, 2015, 10(6):845-858.
[13] 李玲玲, 朱珊丽, 李文姝, 等. EB病毒核蛋白-1 B细胞表位的预测及其同源性分析[J]. 生物医学工程学杂志, 2011, 28(2):371-375.
LI L L, ZHU S L, LI W S, et al. Prediction and research on homology of B-cell epitopes of Epstein-Barr virus nuclear antigen-1[J]. Journal of Biomedical Engineering, 2011, 28(2):371-375. (in Chinese)
[14] SIVALINGAM G N, SHEPHERD A J. An analysis of B-cell epitope discontinuity[J]. Mol Immunol, 2012, 51(3-4):304-309.
[15] BLAZIE S M, BABB C, WILKY H, et al. Comparative RNA-Seq analysis reveals pervasive tissue-specific alternative polyadenylation in Caenorhabditis elegans intestine and muscles[J]. BMC Biol, 2015, 13:4.
[16] LIU G H, XU M J, CHANG Q C, et al. De novo transcriptomic analysis of the female and male adults of the blood fluke Schistosoma turkestanicum[J]. Parasit Vectors, 2016, 9:143.
[17] 成文敏, 潘伟荣, 尹俊芳, 等. 版纳微型猪近交系DLDH基因编码区序列扩增及生物信息学分析[J]. 畜牧兽医学报, 2015, 46(10):1706-1712.
CHENG W M, PAN W R, YIN J F, et al. Amplification and bioinformatics analysis of coding region of the DLDH gene in Banna Mini-pig inbred line (BMI)[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(10):1706-1712. (in Chinese)
[18] TRITTEN L, CLARKE D, TIMMINS S, et al. Dirofilaria immitis exhibits sex-and stage-specific differences in excretory/secretory miRNA and protein profiles[J]. Vet Parasitol, 2016, 232:1-7.
[19] EBERLE R, BRATTIG N W, TRUSCH M, et al. Isolation, identification and functional profile of excretory-secretory peptides from Onchocerca ochengi[J]. Acta Trop, 2015, 142:156-166.
[20] CAO X D, FU Z Q, ZHANG M, et al. iTRAQ-based comparative proteomic analysis of excretory-secretory proteins of schistosomula and adult worms of Schistosoma japonicum[J]. J Proteomics, 2016, 138:30-39.
[21] CAO X D, FU Z Q, ZHANG M, et al. Excretory/secretory proteome of 14-day schistosomula, Schistosoma japonicum[J]. J Proteomics, 2016, 130:221-230.

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

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 14

Recommended Articles

Metrics

Comments

[1]	BO Luqi, WANG Xueying, HOU Jiaming, ZHANG Li, ZHANG Jianhui, RUAN Rulin, ZHANG Bohan, WANG Shuang, SONG Mingxin, ZHANG Ziqun. Interference of Ts-ODC Gene of Trichinella spiralis by siRNA-757 Effect on Its Acid Resistance [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2114-2125.
[2]	WANG Bingnan, SUN Xue, JIA Hongguo, TAN Chun, MEI Sipeng, ZHOU Rongqiong. Prokaryotic Expression and Tissue Localization of C-type Lectin 4 from Toxocara canis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1641-1651.
[3]	LIANG Gaoxing, RONG Shiqi, WANG Junwei, LI Yuan, YANG Xin, ZHAO Guanghui, SONG Junke. Multi-locus Gene Sequence Analysis of Female Haemonchus contortus with Morphological Differences in Vulval Flap from Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3140-3148.
[4]	LIAO Shuyi, CHU Wen, XU Lixin, SONG Xiaokai, LI Xiangrui, YAN Ruofeng. Effect of Trichinella spiralis Recombinant Dipeptidyl-peptidase 1 on Proliferation, Phagocytosis, and Apoptosis of Rat PBMCs in vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1362-1370.
[5]	CHEN Shaoji, LI Fang, YIN Shasha, JIANG Aiyun, ZHOU Rongqiong. Expression and Chitin-Binding Feature of TcCPG1 of Toxocara canis [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(2): 356-364.
[6]	LUO Yongli, JIANG Aiyun, HU Zhigang, LI Xianglan, YANG Xiaodi, KUANG Ceyan, ZHOU Rongqiong. Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 390-397.
[7]	WU Yiran, GU Xiaobin, XIE Yue, YANG Guangyou. Prokaryotic Expression and Primary Evaluation of Antibacterial Activity of Antibacterial Factor in Baylisascaris schroederi [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 151-158.
[8]	ZHOU Li-na, HU Meng-juan, SUN Wei, LI Xiang-rui, XU Li-xin, SONG Xiao-kai, YAN Ruo-feng. Gene Cloning and Biological Characteristics of Lipase Gene from Haemonchus contortus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 580-587.
[9]	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.
[10]	WU Ling-yan, WANG Yu-jian, WEN Yu-ling, YAN Ruo-feng, XU Li-xin, SONG Xiao-kai, LI Xiang-rui. Cloning, Expression and Function Analysis of NADH: Ubiquinone Oxidoreductase Domain Containing Protein in Haemonchus contortus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(4): 722-730.
[11]	ZHENG You-le，WANG Cheng-dong，WANG Tao. Research Advances on Chemosensory Behaviors of Nematodes [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(5): 857-863.
[12]	GU Xiao-bin，ZHU Jun-yang，WANG Bao-jian，ZHENG Jing，YANG Guang-you，WANG Tao，LAI Wei-min. Genetic Variation of Heterakis gallinarum in Sichuan based on the Nuclear Ribosomal DNA Internal Transcribed Spacer Regions (ITS1/2) [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 796-804.
[13]	WANG Bao-jian，ZHU Jun-yang，JIAN Ke-ling，YANG Guang-you，WANG Tao，LAI Wei-min，ZHAO Xi-bin，LIAO Xiao-xia，GU Xiao-bin. Population Genetic Polymorphisms of Heterakis gallinarum in Sichuan Base on Mitochondrial 12S Gene [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(12): 2264-2272.
[14]	MA Guang-xu，ZHU Hong-hong，HU Ling，LI Wei-lin，LIN Jie，ZHOU Rong-qiong. Differential Expression of Gender-related Genes Tc-vit，Tc-iom，Tc-tpk，Tc-stp in the Germ-line Tissues of Toxocara canis [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(3): 461-466.