甲型H1N1流感病毒毒株的分子特征及功能分析预测

发布时间：2018-01-12 05:30

本文关键词：甲型H1N1流感病毒毒株的分子特征及功能分析预测　出处：《山东大学》2010年硕士论文　论文类型：学位论文

【摘要】： 2009年4月,起源于墨西哥的新型流感引起人们一片恐慌,在短短几周之内,新型流感病毒就迅速地蔓延到了全世界,2009年6月11日,世界卫生组织将全球流感大流行警告级别提升至最高等级第六级,中国乃至山东省都是此次流感的重灾区,影响人们的正常工作生活。此次爆发的流感病毒为一类新型流感病毒,本论文利用结构生物信息学的方法系统分析了山东省毒株的分子特征,并对其功能进行了分析预测。通过分子发育分析,山东省毒株为血凝素(HA)1型,神经氨酸酶(NA)1型毒株。毒株的基因组来源人、猪、禽流感的遗传物质,是一个四重排甲型流感病毒。其HA、NP和NS基因来源于H1N1亚型经典猪流感毒株,NA和MP基因来源于H1N1亚型禽源猪流感毒株。PA和PB2基因来源于不同亚型的禽流感病毒,PBl基因来源于季节性H3N2亚型人流感病毒。虽然新甲型H1N1流感病毒8个基因片段所形成的进化树各不相同,但仍然存在一些共性：4株新甲型H1N1流行病毒株A/Mexico/4486/2009;A/California/04/2009; A/Beijing/3/2009 (H1N1)和A/Shandong/1/2009都聚在一起,形成独立的进化支,并且以高Bootstrap值支持。说明09年新甲型H1N1流感病毒有共同的进化祖先,H1N1流感病毒在流行扩散过程中没有显著变异。对其血凝素蛋白的结构、表面电势分析及分子对接分析表明该甲型H1N1流感病毒的受体结合位点与典型的人流感病毒的氨基酸完全一致,这意味着新型甲型H1N1流感病毒蛋白的受体结合特异性完全符合感染人类的要求,具备人传播人的分子基础的。这也在一定程度上解释了甲型此次H1N1流感病毒只在人与人之间传播,极少见人与猪之间传播的现象。通过该毒株HA蛋白的糖基化、脂化位点分析,发现将毒株的HA蛋白的糖基化位点仅有27位,28位,40位,104位,286位,304位,及498位较相关其它季节性流感病毒变少,脂化位点与二硫键位点未发生变化。山东省毒株具有较少糖基化位点,这暗示该甲型流感病毒应具有较强的致病力。通过基于序列与基于结构的抗原决定簇分析,发现山东省毒株的HA蛋白的抗原决定簇主要位于蛋白的头部,相对澳大利亚毒株,抗原决定簇Cal区222位发生突变(Lys→Arg),相对于2009年美洲区域的毒株未发生明显变化,因此其于世卫组织提供的毒株生产的相关疫苗应该是有效的。通过对NA蛋白结构的预测分析,表明相对于其它季节性流感的NA,此次NA突变的氨基酸位点主要位于活性中心以外的分子表面,未发生NA酶的活性中心,所以针对NA蛋白的相关药物应该是有效的。本文通过对山东省毒株的基因序列、蛋白质序列、蛋白质结构、分子表面性质、分子对接分析、抗原决定簇、药物作用靶位的快速分析,初步建立了快速定型、分析、预测的结构生物信息学方法,该对甲型流感病毒的防控应该具有借鉴与参考意见,特别是对当前山东省当前的疾病防控的基础与形势,本文建立的方法应该会使得相应防控技术由依靠经验转化理性化,特别是随着新一代低成本高通量测序技术的发展,相信结构生物信息学方法将会成为主流技术指导新形势下大流行疾病的防控。
[Abstract]:In April 2009, the new flu originated in Mexico caused people to panic, in a few weeks, the new swine flu virus is spreading rapidly around the world, in June 11, 2009, the WHO will be a global influenza pandemic alert level raised to the highest level sixth, Chinese and Shandong province are the hardest hit by the flu, affecting people's normal working life.
The outbreak of the influenza virus is a new type of influenza virus, method of system by making use of the structural bioinformatics analysis of the molecular characteristics of strains in Shandong Province, and analyzed its function prediction. By molecular phylogenetic analysis, Shandong province strain hemagglutinin (HA) type 1, neuraminidase (NA) type 1 strains. The genome of the strains from human, swine and avian influenza genetic material, is a rearrangement of four influenza A virus. The HA, NP and NS gene from subtype H1N1 of classical swine flu strain, NA source and MP gene from avian H1N1 Subtype Swine influenza virus strain.PA and PB2 gene from different subtypes avian influenza virus, PBl gene derived from seasonal human influenza virus subtype H3N2. Although the phylogenetic tree formed by the new H1N1 influenza virus 8 gene fragments of each are not identical, but there are still some similarities: 4 strains of new influenza H1N1 epidemic strains A/Mexico/4486/200 9; A/California/04/2009; A/Beijing/3/2009 (H1N1) and A/Shandong/1/2009 were clustered together, forming an evolutionary independent branch, and support with high Bootstrap value. 09 years of the new H1N1 influenza virus have a common evolutionary ancestor, H1N1 influenza virus has no significant variation in the popular diffusion process. Structure of the hemagglutinin protein, surface potential analysis and molecular docking analysis showed that the H1N1 influenza virus receptor binding sites of amino acids and typical human influenza virus completely consistent, which means that the new H1N1 influenza virus protein receptor binding specificity in full compliance with the requirements of human infection, and has the molecular basis of human communication. This partly explains the H1N1 influenza virus only spread between people, but seldom spread between human and swine.
The glycosylation of the strains of HA protein, lipid analysis sites, found that the glycosylation site strain HA protein only 27, 28, 40, 104, 286, 304, and 498 than other seasonal influenza viruses become less lipid sites and two sulfur the key points are not changed. Shandong province strains have fewer glycosylation sites, suggesting that the influenza virus with strong pathogenicity. Based on sequence and structure based antigen determinants analysis, found that the epitopes of HA protein in Shandong province were mainly located in the head of Australia protein, virus antigen determinant Cal 222 mutations (Lys, Arg), compared with the 2009 American regional strains did not change significantly, so it provides from who strains vaccine should be effective. Through the prediction of NA protein structure analysis showed that, compared with the For other seasonal influenza NA, the amino acid site of NA mutation is mainly located outside the surface of the active center, and there is no active center of NA enzyme. Therefore, NA related drugs should be effective.
The gene sequence of Shandong strain protein sequence, protein structure, molecular surface properties, molecular docking analysis, antigenic determinant, rapid analysis of target drugs, initially established a fast setting, structure analysis, bioinformatics method, the prevention and control of influenza A virus should have reference opinions, especially based on the current situation in Shandong province and the disease prevention and control, this method should make the corresponding prevention and control technology of rational transformation by relying on experience, especially with the development of a new generation of low cost and high throughput sequencing technology, believe that structural bioinformatics methods will become the mainstream technology of pandemic diseases prevention and control guidance under the new situation.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R373

【引证文献】