摘要
目的获取并分析ORFV AH-F10株VIR基因序列及预测其编码蛋白的生物信息学特点。方法利用实验室保存的羊口疮AH-F10株,设计VIR基因引物并进行PCR扩增、克隆及序列测定,同时利用生物信息学方法对其编码的蛋白的理化性质、二级结构、三级结构、信号肽、磷酸化位点、跨膜结构域以及线性细胞表位进行预测。结果 AH-F10-VIR基因长552 bp,编码183个氨基酸,与Nantou株的VIR基因同源性最高,核苷酸同源性高达99.6%,氨基酸同源性为100.0%。生物信息学分析结果显示编码的蛋白相对分子量为19.88 kDa,等电点为4.83,为亲水性蛋白;α-螺旋、β-转角、无规则卷曲和延伸链分别占36.07%、3.83%、43.17%和16.94%;三级结构预测显示VIR蛋白存在较多的α-螺旋与无规则卷曲,同二级结构预测结果相符;含有17个磷酸化位点,无信号肽和跨膜结构区域,有15个潜在的B细胞优势表位,4个CTL细胞表位以及5个Th细胞表位。结论成功克隆了羊口疮安徽株VIR基因并预测了VIR蛋白的生物信息学相关信息,为进一步研究VIR蛋白奠定了基础。
Objective To obtain and analyze the VIR gene sequence of ORFV AH-F10 strain and predict the bioinformatics characteristics of its encoded protein. Methods Using the laboratory-preserved AH-F10 strain of ORFV, the VIR gene's primer was designed for PCR, cloning and sequencing. Bioinformatics methods were used to predict the physicochemical properties, secondary structure, tertiary structure, signal peptide, phosphorylation site, transmembrane domain and linear cell epitope of the encoded protein. Results The AH-F10-VIR gene was 552 bp in length and encoded 183 amino acids, the homology of which with the VIR gene of Nantou strain was the highest;the nucleotide homology was up to 99.6%, and amino acid homology up to 100.0%. Bioinformatics analysis showed that the encoded protein was a hydrophilic protein with a relative molecular weight of 19.88 kDa and an isoelectric point of 4.83. The α-helices, β-turns, random coils, and extended chains in secondary structure accounted for 36.07%, 3.83%, 43.17%, and 16.94%, respectively;tertiary structure prediction showed that there were more α-helices and random coils in VIR protein, which was consistent with the prediction of secondary structure. It contained 17 phosphorylation sites, with no signal peptide and transmembrane in the structural region, and there were 15 potential B-cell dominant epitopes, 4 CTL cell epitopes and 5 Th cell epitopes. Conclusion The VIR gene of Anhui strain was successfully cloned and the bioinformatics information of its protein was predicted, which provided a basis for further study.
作者
杨侃侃
俞赵荣
张谦
张高峰
王元红
张学琪
鲁智敏
刘自敏
彭开松
王勇
李永东
YANG Kankan;YU Zhaorong;ZHANG Qian;ZHANG Gaofeng;WANG Yuanhong ZHANG Xueqi;LU Zhimin;LIU Zimin;PENG Kaisong;WANG Yong;LI Yongdong(College of Animal Science and Technology,Anhui Agricultural University,Hefei,Anhui 230036,China)
出处
《中国微生态学杂志》
CAS
CSCD
2018年第12期1365-1370,共6页
Chinese Journal of Microecology
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创新创业训练计划项目
