猫泛白细胞减少症病毒北京株NS1基因克隆与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.12.003

摘要/Abstract

摘要：

为分析猫泛白细胞减少症病毒（FPV）北京株（FPV-BJ 05株）NS1基因的分子特征及其编码蛋白的生物学功能，本研究对FPV-BJ 05株NS1基因进行PCR扩增、克隆及序列测定。应用生物信息学软件分析FPV-BJ 05株与GenBank上登录的11株FPV参考株NS1基因的同源性，并预测NS1蛋白理化性质、信号肽、跨膜结构、B细胞抗原表位、磷酸化位点、亚细胞定位及蛋白结构与功能、高级结构等。结果显示，NS1基因全长2 007 bp，编码668个氨基酸，且与其他FPV分离株NS1基因核苷酸序列同源性为98.8%~99.3%，氨基酸序列同源性为97.9%~98.8%。系统进化树分析结果显示，FPV-BJ 05株NS1蛋白与GenBank上登录的3株FPV参考株处于同一大分支，但由于氨基酸的突变导致其属于独立的一小分支。NS1蛋白既无信号肽也无跨膜结构域，属于非分泌型的疏水性蛋白。B细胞抗原表位预测结果显示，NS1蛋白柔韧性较好，抗原性及表面可及性较高，预测该蛋白含有13个优势抗原位点。修饰结构预测表明，NS1蛋白含有62个潜在磷酸化位点，27个O-糖基化位点和2个N-糖基化位点。亚细胞定位结果显示，NS1蛋白在细胞质和细胞核的概率较高，分别为69.6%和17.4%。NS1蛋白二级结构中α-螺旋、延伸链、无规则卷曲和β-转角分别占39.37%、15.42%、39.37%和5.84%。应用在线软件SWISS-Modle对NS1蛋白进行建模，预测其三级结构，结果发现NS1蛋白主要以α-螺旋为主。本试验结果将为北京地区FPV免疫诊断及核酸疫苗研究提供理论依据。

关键词: 猫泛白细胞减少症病毒(FPV); NS1基因; 生物信息学; 蛋白结构预测

Abstract:

In order to study the molecular characteristics and biological function of NS1 gene of feline parvovirus (FPV),the NS1 gene of FPV-BJ 05 strain was amplified,cloned and sequenced.Nucleotide and amino acid homology analysis,signal peptide,transmembrane domain,B-cell preponderant epitope,phosphorylation site subcellular localization and protein structure and fuction,advanced structure of NS1 gene were predicted using bioinformatic softwares.The results showed that the length of NS1 gene was 2 007 bp,encoding 668 amino acids.The NS1 gene of FPV-BJ 05 shared 98.8% to 99.3% nucleotide homology with that of the other FPV strains,and the amino acid homology was 97.9% to 98.8%.Phylogenetic tree analysis showed that the NS1 protein of FPV-BJ 05 strain was classified as the same major branch with 3 FPV reference strains registered in GenBank,but it was a small independent branch due to amino acid mutation.The NS1 protein had neither a signal peptide nor a transmembrane domain,which was a non-secretory hydrophobic protein.The prediction results of B cell antigen parameters showed that NS1 protein had good flexibility,high antigenicity and surface accessibility,and it was predicted that the protein contained 13 dominant antigenic sites.62 phosphorylation sites,27 O-glycosylation sites and 2 N-glycosylation sites were found in NS1 protein.The result of subcellular localization showed that the probability of NS1 protein in the cytoplasm and nucleus was higher,which were 69.6% and 17.4% respectively.The prediction of secondary structure revealed that α-helix (39.37%),extended chain (15.42%),random coil (39.37%) and β-turn (5.84%),and the tertiary structure of NS1 protein was curved spiral structure.This study provided theoretical basis for immunologic diagnosis and further research of nucleic acid vaccine of FPV in Beijing.

Key words: feline panleukopenia virus (FPV); NS1 gene; bioinformatics; protein structure prediction

中图分类号:

S852.65⁺9.2

刘琪, 史利军, 袁维峰, 梁瑞英, 李金祥, 崔尚金. 猫泛白细胞减少症病毒北京株NS1基因克隆与生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(12): 3327-3336.

LIU Qi, SHI Lijun, YUAN Weifeng, LIANG Ruiying, LI Jinxiang, CUI Shangjin. Cloning and Bioinformatic Analysis of NS1 Gene of Feline Parvovirus in Beijing[J]. , 2018, 45(12): 3327-3336.

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