山羊痘病毒A11R基因的克隆及生物信息学分析

发布时间：2018-12-12 08:31

【摘要】：为了分析绵羊痘病毒A11R蛋白质的分子特征,提取了山羊痘病毒古浪分离株(GL)的基因组DNA,设计引物进行PCR扩增,将PCR产物连接到p GEM-T Easy载体后转化至大肠杆菌DH5α感受态细胞,筛选阳性克隆进行序列测定,利用生物信息学软件对A11R基因序列进行预测分析。结果显示,A11R基因序列由954个核苷酸组成的开放阅读框,编码317个氨基酸残基组成的多肽,蛋白质分子质量的理论值为36 091.7,理论等电点为5.14。A11R蛋白质的二级结构中,α螺旋占29.02%,β折叠占10.41%,其余60.57%为无规则卷曲。多序列比对分析显示,不同羊痘病毒分离株A11R序列高度保守,同源性在98%以上。进化树分析结果显示,GL株与NK株、TU株以及SA株在一个分支,说明它们之间具有较近的亲缘关系,上述研究结果为进一步研究A11R蛋白质的生物学功能和羊痘病毒早期蛋白质分子间相互作用奠定了基础。
[Abstract]:In order to analyze the molecular characteristics of sheep pox virus A11R protein, the genomic DNA, design primers of Goat pox virus Gourang isolate (GL) were extracted and amplified by PCR. The PCR product was ligated to p GEM-T Easy vector and transformed into Escherichia coli DH5 伪 competent cells. The positive clones were screened and sequenced. The A11R gene sequence was predicted and analyzed by bioinformatics software. The results showed that the A11R gene sequence was composed of 954 nucleotides and encoded 317 amino acid residues. The theoretical value of protein molecular weight was 36,091.7. The theoretical isoelectric point is the secondary structure of 5.14.A11R protein, 伪 helix is 29.02, 尾 folding is 10.41, the rest 60.57% is irregular crimp. Multiple sequence alignment analysis showed that the A11R sequence of different sheep pox virus isolates was highly conserved, and the homology was over 98%. The results of phylogenetic tree analysis showed that the GL strain was closely related to NK strain, TU strain and SA strain. These results provide a basis for further study on the biological function of A11R protein and the intermolecular interaction between the early proteins of sheep pox virus.
【作者单位】：中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室/农业部畜禽病毒学重点开放实验室;吉林农业大学动物科学技术学院;西北民族大学生命科学技术学院;
【基金】：国家质检总局公益性行业科研专项(201310093) 国家自然科学基金项目(31201892) 甘肃省自然科学基金项目(1208RJZA101) 国家“863”计划项目(2012AA101304)
【分类号】：S852.65

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