Auperin D D, Esposito J J, Lange J V, Bauer S P, Knight J, Sasso D R, McCormick J B
Center for Infectious Diseases, Centers for Disease Control, Atlanta, GA 30333.
Virus Res. 1988 Feb;9(2-3):233-48. doi: 10.1016/0168-1702(88)90033-0.
A cloned cDNA (1.65 kb) containing the complete glycoprotein gene of the Josiah strain of Lassa virus was inserted into the thymidine kinase (TK) gene of the New York Board of Health (WYETH) strain of vaccinia virus. The Lassa virus glycoprotein precursor, GPC, and the posttranslational cleavage products, G1 and G2, were shown by Western blot analysis to be properly expressed in cells infected with the recombinant virus. Northern blot hybridization of total cytoplasmic RNA extracted from recombinant virus infected cells demonstrated the presence of RNA transcripts of appropriate size considering the site of transcription initiation from the vaccinia P7.5 promoter, the size of the Lassa glycoprotein gene, and the presumed location of the transcription terminator in the vaccinia thymidine kinase gene. All guinea pigs vaccinated with the recombinant virus survived a lethal challenge infection with Lassa virus, whereas 80% of control animals died. The vaccinated guinea pigs did, however, develop transient, low-grade, fevers and detectable viremias following infection with Lassa virus, indicating that protection was not complete.
将一个含有拉沙病毒乔赛亚株完整糖蛋白基因的克隆cDNA（1.65 kb）插入痘苗病毒纽约卫生局（惠氏）株的胸苷激酶（TK）基因中。通过蛋白质印迹分析表明，拉沙病毒糖蛋白前体GPC以及翻译后切割产物G1和G2在感染重组病毒的细胞中得到了正确表达。从感染重组病毒的细胞中提取的总细胞质RNA的Northern印迹杂交显示，考虑到从痘苗P7.5启动子的转录起始位点、拉沙糖蛋白基因的大小以及痘苗胸苷激酶基因中转录终止子的假定位置，存在大小合适的RNA转录本。所有接种重组病毒的豚鼠在接受拉沙病毒致死性攻击感染后存活下来，而80%的对照动物死亡。然而，接种疫苗的豚鼠在感染拉沙病毒后确实出现了短暂的低热和可检测到的病毒血症，这表明保护并不完全。
