Yin Jiechao, Ren Xiaofeng, Tian Zhijun, Li Yijing
Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, 150030 Harbin, PR China.
Biologicals. 2007 Mar;35(1):55-61. doi: 10.1016/j.biologicals.2006.02.001. Epub 2006 May 30.
Two severe porcine infectious diseases, pseudorabies (PR) and transmissible gastroenteritis (TGE) caused by pseudorabies virus (PRV) and transmissible gastroenteritis virus (TGEV) respectively often result in serious economic loss in animal husbandry worldwide. Vaccination is the important prevention means against both infections. To achieve a PRV genome-based virus live vector, aiming at further TGEV/PRV bivalent vaccine development, a recombinant plasmid pUG was constructed via inserting partial PK and full-length gG genes of PRV strain Bartha K-61 amplified into pUC119 vector. In parallel, another recombinant pHS was generated by introducing a fragment designated S1 encoding the major antigen sites of S gene from TGEV strain TH-98 into a prokaryotic expression vector pP(RO)EX HTc. The SV40 polyA sequence was then inserted into the downstream of S1 fragment of pHS. The continuous region containing S1fragment, SV40 polyA and four single restriction enzyme sites digested from pHS was subcloned into the downstream of gG promoter of pUG. In addition, a LacZ reporter gene was introduced into the universal transfer vector named pUGS-LacZ. Subsequently, a PRV genome-based virus live vector was generated via homologous recombination. The functionally effective vector was purified and partially characterized. Moreover, the potential advantages of this system are discussed.
两种严重的猪传染病，分别由伪狂犬病病毒（PRV）引起的伪狂犬病（PR）和由传染性胃肠炎病毒（TGEV）引起的传染性胃肠炎（TGE），常常在全球畜牧业中造成严重的经济损失。疫苗接种是预防这两种感染的重要手段。为了构建基于PRV基因组的病毒活载体，旨在进一步开发TGEV/PRV二价疫苗，通过将扩增的PRV株Bartha K-61的部分PK基因和全长gG基因插入pUC119载体，构建了重组质粒pUG。同时，通过将来自TGEV株TH-98的编码S基因主要抗原位点的片段S1引入原核表达载体pP(RO)EX HTc，产生了另一种重组质粒pHS。然后将SV40 polyA序列插入pHS的S1片段下游。将包含S1片段、SV40 polyA和从pHS消化得到的四个单限制酶位点的连续区域亚克隆到pUG的gG启动子下游。此外，将LacZ报告基因引入名为pUGS-LacZ的通用转移载体中。随后，通过同源重组产生了基于PRV基因组的病毒活载体。对功能有效的载体进行了纯化和部分特性鉴定。此外，还讨论了该系统的潜在优势。
