猪流行性腹泻病毒的分离鉴定及其M蛋白截段基因编码氨基酸的生物学分析  

Biological analysis of the amino acid encoded by the truncated M gene after isolation and identification of porcine epidemic diarrhea virus

作　　者：蒋梅[1] 王印[1] 杨泽晓[1] 姚雪萍[1] 彭彬[1] 刘波[1] 邬旭龙[1] 次仁群宗 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]西藏那曲班戈县农牧科学技术服务站,西藏那曲852500

出　　处：《中国兽医学报》2013年第2期195-200,共6页Chinese Journal of Veterinary Science

基　　金：教育部<长江学者和创新团队发展计划>创新团队资助项目(IRT0848);四川农业大学双支计划资助项目(00270401)

摘　　要：利用PK-15细胞从四川省某猪场发病死亡的7日龄仔猪肠系膜淋巴结和脱落的肠黏膜材料中分离获得1株病毒,PCR检测证实为猪流行性腹泻病毒,命名为SC-405。对其M截段基因克隆后进行测序,结果显示该基因长664bp,编码221个氨基酸;序列分析并通过NCBI做序列比对,与其他已知的GenBank公布的序列同源性为97%～99%;氨基酸序列有突变,同源性为95%～99%;系统进化树表明该病毒跟序列号为AEQ29504、ACA81419、AEE38481序列的亲缘关系比较近;用DNAStar Protean模块对该基因编码的蛋白进行主要的抗原指数、二级结构、蛋白质骨架柔性、表面可及性等参数进行预测,并在线预测该蛋白的亲水性及跨膜区,确定该截段基因的B细胞抗原优势表位可能分布在Leu12-Asn14、Lys36、Trp100-Thr113、Val196-Asp202和Asp215-Glu217这些区段。A strain of virus,isolated from porcine mesenteric lymph nodes and exfoliated intestine mucous membrane of 7-day-old piglets which were died from disease in a pig farm in Sichuan province was proved to be porcine epidemic diarrhea virus（PEDV）, named PEDV-SC-405 after PCR test. The truncated M gene of PEDV-SC-405 was cloned and sequenced, showing that 221 amino acids were encoded by the gene（664 bp）. The M gene nucleotide sequence shared 97%-99% homology with it＇s counterpart of other PEDV available in GenBank analysed by BLAST,and amino acid sequence shared 95%-99% identity. A phylogenetic analysis based on the amino acid sequences of M gene indicated that the virus has a close relationship with AEQ29504, ACA81419, AEE38481. Furthermore, the antigenicity, secondary structure, backbone flexibility, surface acces- sibility of the truncated M protein were analyzed by DNAStar Protean software. The hydrophilicity and transmembrane domain of protein were predicted. Meanwhile, the dominant epitope of B-cells from the truncated M gene was identified to distribute those sections such as Leu12-Asn14,Lys36, Trp100-Thrll3, Vall 96-Asp202, Asp215-Glu217.

关 键 词：猪流行性腹泻病毒 M基因 系统进化分析 抗原表位 

分 类 号：S852.65[农业科学—基础兽医学]