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作者将韩国分离的乙型脑炎病毒（JEV）野毒株K94P05的糖基化膜前体（PrM）和包膜(E）蛋白的基因插入到高度减毒的宿主范围限定的痘苗病毒MVA株基因组内,获得两个重组病毒vJH6和vJH9,分别由合成的痘病毒启动子Psyn和修饰的痘苗病毒H5启动子控制。JEV野毒株K94P05的核苷酸序列与JEV原型中山株有88％相同,但是K94P05株的E基因内没有痘菌病毒早期终止信号。应用转移载体将目的JEV基因转移至MVA基因组内的缺失位点组成重组病毒并用JEV抗体免疫染色进行检测,结果表明两个重组体都稳定地表达prM和E蛋白。但vJH9的滴度较vJH6高3～5倍。vJH6和vJH9感染的CEF细胞与抗JEV K94P05株E蛋白表位的单克隆抗体NBl的免疫荧光反应强度与K94P05野毒株相似,而非重组MVA感染的细胞则未显示这种染色结果。用JEV多克隆抗体的结果与单克隆抗体相似。
The authors inserted genes for the glycated membrane precursor (PrM) and envelope (E) proteins of South Korean isolate JEV field strain K94P05 into a highly attenuated host-range-defined vaccinia virus MVA strain Within the genome, two recombinant viruses vJH6 and vJH9 were obtained, which were controlled by the synthetic Pox virus promoter Psyn and the modified vaccinia virus H5 promoter, respectively. The nucleotide sequence of the JEV wild-type strain K94P05 was 88% identical to the JEV prototype Zhongshan strain, but there was no early sign of vaccinia virus in the E gene of the K94P05 strain. Using the transfer vector, the target JEV gene was transferred to the deletion site in the MVA genome to form a recombinant virus and tested by immunostaining with JEV antibody. The results showed that both recombinants stably expressed prM and E proteins. However, the titer of vJH9 is 3 to 5 times higher than that of vJH6. The immunofluorescence intensity of vJH6 and vJH9-infected CEF cells against the monoclonal antibody NBl against the E protein epitope of JEV K94P05 strain was similar to that of the K94P05 wild-type strain, whereas the non-recombinant MVA-infected cells did not show this staining result. The result of using JEV polyclonal antibody is similar to that of monoclonal antibody.