牛乳头状瘤病毒2型贵州株L2基因克隆及生物信息学分析

Cloning and Bioinformatic Analysis of L2 Gene of BPV2 from Guizhou Province

关键词:  牛乳头状瘤病毒2型  贵州株  L2基因  生物信息学分析

Key words:  Bovine papillamavirus 2(BPV2)  Guizhou strain  L2 gene  bioinformatics analysis

摘要:

为分析牛乳头状瘤病毒2型贵州株(BPV2-GZ01株)L2基因的分子特征,预测编码蛋白的生物学功能。本试验对BPV2-GZ01株L2基因进行PCR扩增、克隆及序列测定,应用生物信息学相关软件及方法,对BPV2-GZ01株L2基因进行序列分析并对其编码蛋白进行二级结构、三级结构、B细胞表位、保守结构域、跨膜结构域和信号肽预测。结果显示,L2基因PCR扩增产物大小为1 404 bp,编码467个氨基酸；与参考株BPV2株、BPV2-AKS01株、BPV2-AKS01株、BPV13株、BPV1株相应序列核苷酸同源性分别为99.0%、99.7%、99.3%、83.9%和75.1%,氨基酸的同源性分别为98.9%、99.6%、99.4%、89.5%和82.7%；系统进化显示,BPV2-GZ01株L2基因与BPV2-SW01株亲缘关系最近；二级结构以无规则卷曲、β-折叠和α-螺旋区域所占比例较大,预测此蛋白可能存在B细胞优势抗原表位,无跨膜区域,无信号肽区域。本研究结果将为贵州省BPV核酸疫苗研究提供理论依据。

Abstract:

Abstruct:[Objective]In order to study and analyze the L2 gene of Bovine papillamavirus 2 in Guizhou province(BPV2-GZ01),we studied the scad materials of lambs with clinical sore mouth symptom in Guizhou province.[Methods]The L2 gene was amplified,cloned and sequenced using bioinformatic softwares and methods,the secondary structure, tertiary structure,B-cell preponderant epitope,conserved domains analysis,transmembrane domain and signal peptide of L2 gene were predicted.[Results]The results indicated the length of L2 gene was 1 404 bp,encoding 467 amino acids.The L2 gene of BPV2-GZ01 strain shared a nucleotide identities of 99.0%、99.7%、99.3%、83.9% and 75.1%,and an amino acid identities of 98.9%、99.6%、99.4%、89.5% and 82.7% with those of strains BPV2,BPV2-AKS01,BPV2-AKS01,BPV13 and BPV1,respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between BPV2-GZ01 strain and BPV2-SW01.Prediction of the secondary structure of L2 indicated that the random coil,extended strand and alphahelix took a higher percentage.The L2 protein was supposed contain antigen epitopes,and have a transmembrane domains,no signal peptide found.Tertiary structure is curved spiral structure.[Conclusion]These results provided a theoretical basis for further research of nucleic acid vaccine of BPV.