埃博拉病毒包膜糖蛋白的密码子偏爱性分析

发布时间：2017-12-27 17:37

本文关键词：埃博拉病毒包膜糖蛋白的密码子偏爱性分析　出处：《医学研究杂志》2015年03期 　论文类型：期刊论文

【摘要】：目的对从2014年西非埃博拉暴发地区感染者分离的4个埃博拉病毒株基因组包膜糖蛋白编码基因进行密码子偏爱性分析,为埃博拉病毒包膜蛋白基因表达中宿主系统选择和密码子优化提供参考。方法通过Gene Bank搜索近期公布的分离自西非地区的4个埃博拉病毒株全基因组序列,获得编码包膜糖蛋白GP1和GP2的基因序列及蛋白编码序列。采用EMBOSS在线预测埃博拉病毒包膜糖蛋白的密码子偏爱性,并通过与Codon Usage Database中大肠杆菌、酵母及人的密码子使用频率进行比较,筛选最适埃博拉病毒GP蛋白体外表达的宿主物种,并提供相应密码子优化方案。结果埃博拉病毒基因组、GP1和GP2的Nc值均在57左右,CAI值均在0.7左右,表明其密码子偏爱性较均一,且基因的表达水平相对较高;编码GP1蛋白F、H、M、N、W等氨基酸和GP2蛋白D、K、N、Q、Y等氨基酸中不同密码子使用频率有较大差异;GP1和GP2蛋白密码子偏爱性有共同之处也存在明显差异。计算σGP/E.coli、σGP/Yeast、σGP/Human的比值,结果显示GP1中2.0或0.5的数量分别为24、20和19个;GP2中的数量分别为26、30和19,表明埃博拉病毒GP1和GP2蛋白与人的密码子使用频率较为接近。结论以密码子偏爱性分析作为辅助手段,分析出此次流行于西非的埃博拉病毒包膜糖蛋白的密码子偏爱性及最适表达物种及密码子优化方案,为体外表达GP1和GP2蛋白,从而研发埃博拉病毒检测试剂、亚单位疫苗及基因工程抗体提供参考。
[Abstract]:Objective to analyze codon bias in genome envelope glycoprotein coding genes of 4 Ebola virus isolates from West Africa Ebola outbreak area in 2014, so as to provide references for host system selection and codon optimization in Ebola virus envelope protein gene expression. Methods the genome sequences of 4 Ebola strains isolated from West Africa were searched by Gene Bank, and the gene sequences and protein coding sequences encoding envelope glycoproteins GP1 and GP2 were obtained. The codon preference of using EMBOSS online prediction of Ebola virus glycoprotein, and with the Codon Usage Database in Escherichia coli, yeast and human codon use frequency are compared, the optimum screening of Ebola virus GP protein expression in vitro of the host species, and to provide the corresponding codon optimization scheme. The results of the Ebola virus genome, GP1 and GP2 Nc values were about 57, CAI value was about 0.7, indicated that the codon preference was uniform, and the expression level of the gene encoding GP1 protein is relatively high; F, H, M, N, W, D, amino acid and GP2 protein K, N, Q Y and other amino acids in different codon usage is different; GP1 and GP2 protein codon preference common also have obvious difference. The ratios of sigma GP/E.coli, sigma GP/Yeast and sigma GP/Human were calculated. The results showed that the number of 2 or 0.5 in GP1 was 24, 20 and 19, respectively. The number of GP2 was 26, 30 and 19 respectively, indicating that the GP1 and GP2 protein of Ebola virus were close to the frequency of human codon usage. Conclusion the codon preference analysis as auxiliary means, analysis of the codon preference of the epidemic in West Africa Ebola virus glycoprotein and the optimal expression of species and the codon optimization scheme for in vitro expression of GP1 and GP2 protein, thus developed Ebola virus detection reagents, subunit vaccine and gene engineering antibody reference.
【作者单位】：中国医学科学院/北京协和医学院医学生物学研究所、云南省重大传染病疫苗研发重点实验室;
【基金】：国家自然科学基金资助项目(面上项目)(81171946) 云南省科技厅社会发展科技计划项目(2011CA016) 云南省自然科学基金资助项目(2009ZC187M,2012FB188) 中国医学科学院医学生物学研究所重大专项基金资助项目(2014MB05ZD)
【分类号】：R373
【正文快照】： 据世界卫生组织(World Health Organization,WHO)报道,埃博拉病毒(Ebolavirus)疫情在西非快速暴发,截止2014年7月30日已有767例死亡病例,最新数据似乎表明疫情正在失控[1]。目前,为防止埃博拉病毒传入中国,边境检疫部门已加强相关检查。因此,迅速研究针对埃博拉病毒的检测试剂

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