[目的]了解上海地区鸽Ⅰ型副黏病毒(PPMV-1)流行毒株F基因遗传变异情况，分析F蛋白生物信息学特征，为PPMV-1的防控提供技术支持。[方法]应用RT-PCR方法扩增分离株F基因，测序并进行遗传进化分析，并通过生物信息学在线软件对F蛋白结构功能进行预测分析。[结果]分离株F基因编码区全长为1 662 bp，编码553个氨基酸，F蛋白裂解位点序列为112R-R-Q-K-R-F117，符合强毒株序列结构特征。系统进化树分析结果显示，分离株属于新城疫病毒(NDV)Class Ⅱ群Ⅵ。2。1。1。2。2亚型，与疫苗株La Sota和Mukteswar处于不同进化分支，与国内分离株Pi/SH/CH/0617/2013和pigeon/Ningxia/2068/2016同属Ⅵ。2。1。1。2。2分支。生物信息学分析结果表明，F蛋白为亲水性蛋白，分子质量约为59。03 ku，理论等电点(PI)为7。87，半衰期为30 h，不稳定系数为35。06，脂肪系数为109。73。F蛋白主要分布在细胞质膜和内质网中，具有跨膜结构和信号肽，包含6个潜在的N-糖基化位点、13个半胱氨酸残基、103个O-糖基化位点，以及67个磷酸化位点。二级结构中α-螺旋、无规则卷曲、延伸链、β-转角分别占46。11％、31。46％、18。08％及4。34％，三级结构预测结果与二级结构相一致。B细胞抗原位点预测结果显示，F蛋白含有11个抗原表位(氨基酸数目≥7)。[结论]本研究分离获得的PPMV-1分离株属于NDV Class Ⅱ群Ⅵ。2。1。1。2。2亚型，其F蛋白具有较好的抗原表位，可作为PPMV-1疫苗研制和抗病毒治疗的靶蛋白。研究结果为有效防控PPMV-1感染提供了一定的参考依据，同时也为进一步研发新型疫苗奠定了基础。
Phylogenetic and Bioinformatics Analysis of F Gene of Pigeon Paramyxovirus Type Ⅰ from Shanghai Area
[Objective]The purpose of this study was to understand the genetic variation of F gene of Pigeon paramyxovirus type Ⅰ(PPMV-1)epidemic strain in Shanghai area,analyze the bioinformatic characteristics of F protein,and provide technical support for the prevention and control of PPMV-1.[Method]F gene of the isolate was amplified by RT-PCR,and sequencing and genetic evolution analysis were performed.The structure and function of F protein were predicted by online bioinformatics software.[Result]The total length of the F gene coding region of the isolate was 1 662 bp,encoding 553 amino acids,and the F protein cleavage site sequence was 112R-R-Q-K-R-F117,which was consistent with the sequence structure of the virulent strain.Phylogenetic tree analysis showed that the isolate belonged to Class Ⅱ group Ⅵ.2.1.1.2.2 subtype of Newcastle disease virus(NDV).The isolate was in a different evolutionary branch from the vaccine strains La Sota and Mukteswar,and belonged to the same branch Ⅵ.2.1.1.2.2 with the domestic isolates Pi/SH/CH/0617/2013 and pigeon/Ningxia/2068/2016.The results of bioinformatics analysis showed that F protein was a hydrophilic protein with molecular mass of 59.03 ku,theoretical isoelectric point(pI)of 7.87,half-life period of 30 h,instability coefficient of 35.06 and fat coefficient of 109.73.F protein was mainly distributed in the plasma membrane and endoplasmic reticulum of the cell,and had a transmembrane structure and signaling peptides,including 6 potential N-glycosylation sites,13 cysteine residues,103 O-glycosylation sites,and 67 phosphorylation sites.In the secondary structure,alpha helix,random coil,extended chain and beta turn account for 46.11％,31.46％,18.08％and 4.34％,respectively.The predicted result of tertiary structure was consistent with the secondary structure.B-cell antigen site prediction showed that F protein contained 11 epitopes(amino acid number ≥7).[Conclusion]The PPMV-1 isolate belonged to NDV Class Ⅱ group Ⅵ.2.1.1.2.2 subtype of NDV,and its F protein had a good epitope,which could be used as the target protein for PPMV-1 vaccine development and antiviral therapy.The results provided a certain reference for effective prevention and control of PPMV-1 infection,and also laid a foundation for further research and development of new vaccines.
Pigeon paramyxovirus type ⅠF genephylogeneticbioinformatics
万方数据
维普
