Expression of Pseudorabies Virus gE Core Epitopes in Escherichia coli Strain BL21 and Utilization of Indirect PRV gE-ELISA  

作　　者：Guangjun GUO Sufang LU Guanggang QU Feng LI Lin DONG Yanli BI Jinliang WANG Feng WEI Na TANG Chunling ZHANG Zhuang DING Zhiqiang SHEN 

机构地区：[1]Shandong Binzhou Animal Science & Veterinary Medicine Academy [2]College of Veterinary Medicine,Jinlin University [3]Shandong Lvdu Bio-Science & Technology Co. ,Ltd.

出　　处：《Agricultural Biotechnology》2014年第4期39-44,共6页农业生物技术（英文版）

基　　金：Supported by Shandong Provincial Natural Science Foundation of China(ZR2012CQ012);Shandong Provincial Technical Innovation Grant of China(201220916006)

摘　　要：Pseudorabies virus glycoprotein E （PRV gE） has been recognized as a suitable diagnostic antigen for pseudorabies. In order to produce gE antigen in large quantities and at low cost, a gene fragment encoding PRV gE core epitopes was expressed in E. coli BL21 expression system. SDS-PAGE and Western Blotting revealed that the expression product in culture supematant of E. coli BL21 was a recombinant protein, approximately 51.8 Kd. At 5 h post-induction, protein concentration assay showed that the expression product amounted to 1.65 mg/ml, accounting for 24. 17% of total proteins in the culture supematant. An indirect PRV gE-ELISA was established by using the recombinant expression product as a coating antigen. Cross-reactivity assay showed that this antigen was PRV specific. In addition, the assay was consistently reproducible. Comparison of detection results of 240 serum samples between PRV gE-ELISA and a commercially available PRV diagnostic kit showed that there was no significant difference between these two methods （P 〉 0.05 ）.Pseudorabies virus glycoprotein E （PRV gE） has been recognized as a suitable diagnostic antigen for pseudorabies. In order to produce gE antigen in large quantities and at low cost, a gene fragment encoding PRV gE core epitopes was expressed in E. coli BL21 expression system. SDS-PAGE and Western Blotting revealed that the expression product in culture supematant of E. coli BL21 was a recombinant protein, approximately 51.8 Kd. At 5 h post-induction, protein concentration assay showed that the expression product amounted to 1.65 mg/ml, accounting for 24. 17% of total proteins in the culture supematant. An indirect PRV gE-ELISA was established by using the recombinant expression product as a coating antigen. Cross-reactivity assay showed that this antigen was PRV specific. In addition, the assay was consistently reproducible. Comparison of detection results of 240 serum samples between PRV gE-ELISA and a commercially available PRV diagnostic kit showed that there was no significant difference between these two methods （P 〉 0.05 ）.

关 键 词：Pseudorabies virus Glycoprotein E PRV strain SA Gene expression ELISA 

分 类 号：S852.65[农业科学—基础兽医学] S859.84[农业科学—兽医学]