Fraisier Christophe, Arnarson Hallgrimur, Barbezange Cyril, Andrésdŏttir Valgerdur, Carrozza Maria Luisa, De Andrés Damian, Tolari Francesco, Rosati Sergio, Luján Lluis, Pépin Michel, Amorena Beatriz, Harkiss Gordon, Blacklaws Barbara, Suzan-Monti Marie
Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France.
J Virol Methods. 2007 Dec;146(1-2):363-7. doi: 10.1016/j.jviromet.2007.06.015. Epub 2007 Aug 1.
There are very few previous reports of expression of native full-length maedi visna virus (MVV) Env gp150 protein in the literature. Therefore the use of different plasmid and viral expression vectors to obtain full-length gp150 was investigated. A mammalian expression plasmid, pN3-Env, was constructed containing the MVV env gene encoding the precursor protein gp150 Env. The functionality of the recombinant plasmid was tested for expression in HEK293 cells. A recombinant modified vaccinia Ankara virus, MVA-Env, with expression detected in avian cells was also made. The expression of the MVV gp150 Env precursor protein was shown for the first time upon transfection of the eukaryotic HEK293 cells by the pN3-Env plasmid DNA as demonstrated by Western blot analysis. These plasmid or viral expression vectors are of potential use in MVV vaccines.
此前在文献中很少有关于天然全长梅迪-维斯纳病毒（MVV）Env gp150蛋白表达的报道。因此，研究了使用不同的质粒和病毒表达载体来获得全长gp150。构建了一种哺乳动物表达质粒pN3-Env，其包含编码前体蛋白gp150 Env的MVV env基因。测试了重组质粒在HEK293细胞中的表达功能。还制备了一种在禽细胞中检测到表达的重组改良安卡拉痘苗病毒MVA-Env。如蛋白质印迹分析所示，pN3-Env质粒DNA转染真核HEK293细胞后，首次显示了MVV gp150 Env前体蛋白的表达。这些质粒或病毒表达载体在MVV疫苗中具有潜在用途。
