Expression of Pseudorabies Virus gE Core Epitopes in Escherichia coli Strain BL21 and Utilization of Indirect PRV gE-ELISA
Expression of Pseudorabies Virus gE Core Epitopes in Escherichia coli Strain BL21 and Utilization of Indirect PRV gE-ELISA作者机构：Shandong Binzhou Animal Science & Veterinary Medicine Academy College of Veterinary MedicineJinlin University Shandong Lvdu Bio-Science & Technology Co. Ltd.
出 版 物：《Agricultural Biotechnology》 (农业生物技术（英文版）)
年 卷 期：2014年第3卷第4期
页 面：39-44页
学科分类：090603[农学-临床兽医学] 090601[农学-基础兽医学] 09[农学] 0906[农学-兽医学]
基 金：Supported by Shandong Provincial Natural Science Foundation of China(ZR2012CQ012) Shandong Provincial Technical Innovation Grant of China(201220916006)
主 题：Pseudorabies virus Glycoprotein E PRV strain SA Gene expression ELISA
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 ）.
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