摘要
为更好地了解近年来中国暴发的猪流行性腹泻(porcine epidemic diarrhea,PED)疫情,本研究采集荣昌及其周边地区疑似患有PED的猪小肠上皮组织进行RNA的提取,扩增猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)M基因序列,进行序列分析及M蛋白分子特性研究。结果显示,M基因开放阅读框(ORF)长为681bp,编码226个氨基酸。通过DNAMAN软件及系统进化树分析发现,本试验中PEDV与GenBank中参考毒株的M基因氨基酸同源性在96.0%以上,其中与广东毒株CH/SD-M/2012株亲缘关系最为相近,同源性高达96.9%,表明M基因相对比较保守。通过M蛋白同源建模及功能预测发现,M蛋白晶体模型同SARS冠状病毒nsp14-nsp10复合体5c8s.1.A及NAD激酶Ⅰ2i1w.1.B模型序列相似性为27.69%、15.07%;其高级结构中在第110—113、118—125、132—136、140—145、147—152、154—157、160—173位氨基酸处存在7个α螺旋,在第98—100、102—107、135—137、139—145、149—154氨基酸存在5个锌指结构配体,M蛋白为PEDV病毒基因组复制酶的多聚蛋白。
In order to study the porcine epidemic diarrhea（PED）epidemic situation recently,small intestine tissue from piglets suspected porcine epidemic diarrhea virus（PEDV）were collected in this study.The PEDV M gene was amplified using RT-PCR method and the molecular characterization were analysed.The results showed that the ORF of Mgene was 681 bp which encoded 226 amino acids.The percent identity of M gene amino acid between PEDV in this study and reference strains in GenBank were higher than 96.0%,and that with CH/SD-M/2012 strain was highest（96.9%）.The phylogenetic tree based on the Neighbor-Joining（NJ）and Minimum-Evolution（ME）showed that the PEDV in this study was closely related to CH/SD-M/2012 strain which were belonged to the same evolutionary branch.The phylogenetic tree analysis illustrated that M gene of PEDV was relatively conservative.The results of homology modeling analysis found that crystal model of M protein was similar to coronavirus nsp14-nsp10 complex 5c8s.1and NAD kinaseⅠ 2i1 w.1.B which shared 27.69% and 15.07% sequence similarity,respectively.The protein structure prediction analysis found sevenα-helix structures located in 110 to 113,118 to 125,132 to 136,140 to 145,147 to 152,154 to 157and 160 to 173amino acid regions,and five ligand structures of zinc finger located in 98 to 100,102 to 107,135 to 137,139 to 145and 149 to 154,amino acid regions.The protein structure prediction analysis indicated that the M protein might be a polyprotein of viral genome replication enzyme.
作者
毕俊萱
陈奕熹
龙安格
赵茹钰
张涵铱
唐明霞
易华山
马鲜平
BI Jun-xuan CHEN Yi-xi LONG An-ge ZHAO Ru-yu ZHANG Han-yi TANG Ming-xia YI Hua-shan MA Xian-ping(CoLlege of Animal Science, Southwest University, Chongqing 402460, Chin)
出处
《中国畜牧兽医》
CAS
北大核心
2017年第9期2573-2579,共7页
China Animal Husbandry & Veterinary Medicine
基金
西南大学荣昌校区青年基金项目(RC20700421
RC20700925)
关键词
猪流行性腹泻病毒
M基因
基因克隆
序列分析
porcine epidemic diarrhea virus
M gene
gene cloning
sequence analysis
