Rosen E, Stapleton J T, McLinden J
American Biogenetic Sciences, Inc., University of Notre Dame, IN 46556.
Vaccine. 1993;11(7):706-12. doi: 10.1016/0264-410x(93)90253-t.
Recombinant baculoviruses were constructed which contained the hepatitis A virus (HAV) open reading frame (ORF) under the control of the polyhedrin promoter. Northern blot analysis with an HAV-specific oligonucleotide probe demonstrated a single transcript large enough to include the HAV ORF in Spodoptera frugiperda cells infected with these recombinants. Immunoblots revealed a 220 kDa protein representing the HAV polyprotein. In addition, proteins which co-migrated with HAV capsid proteins, and several proteins of intermediate size were present, consistent with processing intermediates. HAV antigen was present in cells infected with the recombinant baculoviruses when assessed by solid-phase radioimmunoassay. This HAV antigen had a buoyant density in caesium chloride gradients similar to HAV empty capsids, and elicited HAV neutralizing antibodies in mice.
构建了重组杆状病毒，其在多角体蛋白启动子的控制下包含甲型肝炎病毒（HAV）开放阅读框（ORF）。用HAV特异性寡核苷酸探针进行的Northern印迹分析表明，在感染这些重组体的草地贪夜蛾细胞中，有一个足够大的单一转录本，足以包含HAV ORF。免疫印迹显示一条220 kDa的蛋白质，代表HAV多蛋白。此外，存在与HAV衣壳蛋白共迁移的蛋白质，以及几种中等大小的蛋白质，这与加工中间体一致。通过固相放射免疫测定评估时，重组杆状病毒感染的细胞中存在HAV抗原。这种HAV抗原在氯化铯梯度中的浮力密度与HAV空衣壳相似，并在小鼠中引发HAV中和抗体。
