Higashihashi N, Arai Y, Enjo T, Horiuchi T, Saeki Y, Sakano K, Sato Y, Takeda K, Takashina S, Takahashi T
Molecular Biology Research Laboratory, Daiichi Pharmaceutical Co., Ltd., Edogawa-ku, Tokyo, Japan.
J Virol Methods. 1991 Nov-Dec;35(2):159-67. doi: 10.1016/0166-0934(91)90131-i.
The Bombyx mori nuclear polyhedrosis virus (BmNPV) in the silkworm was used successfully for mass production of biologically active foreign genes under the control of the polyhedrin promoter. This system was adapted for the production of large amounts of hepatitis B virus surface antigens (HBsAg). The DNA fragments coded for the middle protein, which is composed of the S protein with the pre-S2 region, were cloned, the signal protein gene of beta-IFN was added, and both were inserted into a cloning vector. After co-transfection with wild-type BmNPV, stable recombinant viruses were isolated by the limiting dilution method. Infected silkworm larvae with the recombinants expressed HBsAg at high levels (400-600 micrograms/ml). These products, consisting of two polypeptides with molecular weights of approximately 25,000 (p25) and glycosylated P25 (GP28), were purified as assembled 22-nm particles. We demonstrated that HBsAg from silkworms consists of S protein with 7 amino acids of Pre-S2.
家蚕核型多角体病毒（BmNPV）被成功用于在多角体蛋白启动子控制下大规模生产具有生物活性的外源基因。该系统被用于大量生产乙型肝炎病毒表面抗原（HBsAg）。编码由带有前S2区的S蛋白组成的中蛋白的DNA片段被克隆，添加了β-干扰素的信号蛋白基因，二者都插入到一个克隆载体中。与野生型BmNPV共转染后，通过有限稀释法分离出稳定的重组病毒。用重组病毒感染家蚕幼虫可高水平表达HBsAg（400 - 600微克/毫升）。这些产物由分子量约为25,000的两种多肽（p25）和糖基化的P25（GP28）组成，作为组装好的22纳米颗粒被纯化。我们证明家蚕来源的HBsAg由带有前S2区7个氨基酸的S蛋白组成。
