马疱疹病毒1型gB糖蛋白线性B细胞表位的预测与鉴定

doi:10.16431/j.cnki.1671-7236.2017.05.032

Abstract

Abstract:

The study was aimed to predict B cell epitopes in gB glycoprotein of equine herpesvirus type 1 (EHV-1) with bioinformatics, and select epitopes which had potential diagnostic value. The DNA fragments of gB glycoprotein were predicted by protean of DNAStar software. Screening potential B cell epitopes after parameter comparison, the target B cell epitopes were selected, cloned and expressed. The expressed fusion proteins serviced as an antigen were used to react with equine herpesvirus positive serum to screen and identify antigenic epitopes. The results showed that according to predictive and analysis, the areas of amino acid from 6 to 10, 23 to 32, 53 to 65, 72 to 98, 111 to 120, 152 to 166, 173 to 180 might be gB glycoprotein B cell epitopes. Seven epitopes were successfully cloned into a prokaryotic expression vector, and confirmed by DNA sequencing. After expression and purification, Western blotting was performed to detect the antigen, which could be recognized by equine herpesvirus positive sera. Bioinformatics technology and molecular biology techniques were used to successfully screen five potential B cell epitopes, which provided the foundation for the diagnosis of EHV-1 and design of the epitope vaccine.

Key words: B-cell epitope; antigenic epitope prediction; prokaryotic expression; identification

CLC Number:

S852.65⁺2

LI Jing, LIU Jian-hua, FU Qiang, SONG Huan-tang, FAN Bin, HU Yue, RAN Duo-liang. Prediction and Identification of Linear B-cell Epitopes in gB Glycoprotein of Equine Herpesvirus Type 1[J]. , 2017, 44(5): 1484-1490.

References

[1] 杨夷平,刘建华,冉多良,等.马疱疹病毒1型LAMP检测方法的建立[J].中国畜牧兽医,2014,41(90):35-40.
[2] Mahmoud H Y, Andoh K, Hattori S, et al. Characterization of glycoproteins in equine herpesvirus-1[J]. J Vet Med Sci,2013, 75(10):1317-1321.
[3] Enio M, Maria C L, Elenice M S, et al. Molecular characterization of Brazilian equid herpesvirus type 1 strains based on neuropathogenicity markers[J].Sociedade Brasileira de Microbiologia,2015,46(2):565-570.
[4] Tracy S, Wee M Y, Deborah B, et al. Equid herpesvirus type 1 activates platelets[J].PLoS One,2015,10(14):e0122640.
[5] Bart S, Nikolaus O, Walid A, et al. Comparative analysis of glycoprotein B (gB) of equine herpesvirus type 1 and 4(EHV-1 and EHV-4) in cellular tropism and cell-to-transmission[J]. Viruses, 2015, 3390(10):522-542.
[6] Atanasiu D, Whitbeck J C, Cairns T M, et al. Bimolecular complementation reveals that glycoproteins gB and gH/gL of herpes simplex virus interacts with each other during cell fusion[J]. Proc Natl Acad Sci USA,2007, 104(7):18718-18723.
[7] Turtinen L W, Allen G P, Gen V,et al. Identification of the envelope surface glycoproteins of equine herpesvirus type 1[J]. Virology, 1982, 63(2):481-485.
[8] Connolly S A, Jackson J O, Jaidetzky T S, et al. Fusing structure and function:A structural view of the herpesvirus entry machinery[J]. Microbiol, 2011, 9(5):369-381.
[9] Eubauer A, Beer M, Brandmu E C, et al. Equine herpesvirus 1 mutants devoid of glycoprotein B or M are apathogenic for mice but induce protection against challenge infection[J]. Virology, 1997, 239(1):36-45.
[10] 李娜. 马疱疹病毒1型gB/gD双基因原核表达蛋白免疫效果研究以及新疆部分地区马鼻肺炎血清流行病学调查[D]. 乌鲁木齐:新疆农业大学, 2014.
[11] 时成龙, 侯玉杰, 相文华, 等. 马鼻肺炎病毒主要糖蛋白及其功能[J]. 动物医学展, 2009, 30(12):90-93.
[12] Shan L Z, Yan F, Xiao Y L, et al. Identification of linear B-cell epitopes within tarp of chlamydia trachomatis[J]. Peptide Science, 2014, 20(6):916-922.
[13] 萨姆布鲁克J,拉塞尔D W. 黄培堂译. 分子克隆实验指南[M]. 第3版. 北京:科学技术出版社, 2002.
[14] 李娜,孙建华,石明,等. 新疆部分地区马鼻肺炎的血清学调查[J]. 动物医学进展,2014, 35(9):115-117.
[15] 沙燕,孙建华,杜润慈,等. 新疆伊犁地区部分马病血清学调查[J]. 畜牧与兽医,2016,48(8):91-94.
[16] 马凡舒, 张蕾, 王洋,等. B细胞抗原表位预测方法的研究进展[J]. 中国畜牧兽医, 2016, 43(1):63-67.
[17] 宋帅, 李春玲, 贾爱卿, 等. 抗原表位研究方法进展[J]. 动物医学进展, 2010, 31(12):87-91.
[18] 董娇娇, 刘子钰, 刘春宇,等. 线性B细胞表位预测应用研究[J]. 计算机光盘软件与应用, 2013, 24(7):63-64
[19] 文雪霞, 陈化兰, 熊永忠, 等. 抗原表位鉴定方法的研究进展[J]. 中国畜牧兽医, 2012, 39(7):66-70.
[20] 宋焕堂,孙建华,杜润慈,等. 马疱疹病毒抗体间接ELISA检测方法的建立[J]. 畜牧与兽医,2016,48(6):1-5.

Prediction and Identification of Linear B-cell Epitopes in gB Glycoprotein of Equine Herpesvirus Type 1

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