摘要
试验旨在克隆表达羊口疮病毒安徽株023基因,对该基因进行PCR扩增、亚克隆及表达,通过蛋白表达纯化后进行SDS-PAGE和Western blot鉴定,并利用生物信息学软件预测该基因编码的蛋白的理化性质、二级结构、信号肽、跨膜结构域等。结果显示,023基因全长819 bp,编码272个氨基酸,蛋白大小约为42 ku,主要以包涵体的形式表达且有良好的反应原性。生物信息学分析表明,该蛋白是亲水稳定蛋白,其中α-螺旋(h)占27. 57%,延伸链(e)占17. 28%,β-转角(t)占12. 13%,无规则卷曲(c)占43. 01%,无信号肽区域和跨膜结构域,可能存在30个磷酸化位点,4个B细胞抗原表位,3个CTL表位和3个Th表位。研究结果不仅有助于了解羊口疮病毒023基因的结构与功能,还能为建立快速诊断的检测方法提供参考。
Our study was aimed at cloning and expressing the 023 gene of orf virus(ORFV)Anhui strain,and predicting its biological characteristics.The 023 gene was amplified by PCR,then successfully expressed using a prokaryotic vector.Then,the expressed protein was purified,and identified by SDS-PAGE and Western-blot.In addition,the physicochemical properties,secondary structure,signal peptide,transmembrane domain of the protein was analyzed by bioinformatics software.The results showed that the 023 gene of ORFV Anhui strain was 819 bp in length which could encode 272 amino acids.The size of expressed protein was approximately 42 ku,which was mainly expressed in inclusion body form with good reactogenicity.The results from bioinformatics analysis showed that the protein was a hydrophilic stable protein.There were 27.57%α-helix(h),17.28%extended chain(e),12.13%β-turn(t),43.01%random coli(c).There was no signal peptide region and transmembrane domain.Maybe,it had 30 phosphorylation sites.Moreover,there were four B cell epitopes,three CTL epitopes and three Th epitopes.In conclusion,our study could provide a reference for further study of the structure and function of ORFV 023 gene.
作者
张高峰
杨侃侃
张谦
王元红
俞赵荣
张学琪
鲁智敏
刘自敏
李永东
王勇
ZHANG Gaofeng;YANG Kankan;ZHANG Qian;WANG Yuanhong;YU Zhaorong;ZHANG Xueqi;LU Zhimin;LIU Zimin;LI Yongdong;WANG Yong(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China;Ningbo Municipal Center for Disease Control and Prevention,Ningbo 315010,China)
出处
《浙江农业学报》
CSCD
北大核心
2019年第1期30-38,共9页
Acta Agriculturae Zhejiangensis
基金
国家自然科学基金(31602063)
安徽省自然科学基金(1508085QC60)
安徽农业大学大学生科技创新基金项目
