Motohashi Tomoko, Shimojima Tsukasa, Fukagawa Tatsuo, Maenaka Katsumi, Park Enoch Y
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
Biochem Biophys Res Commun. 2005 Jan 21;326(3):564-9. doi: 10.1016/j.bbrc.2004.11.060.
Silkworm is one of the most attractive hosts for large-scale production of eukaryotic proteins as well as recombinant baculoviruses for gene transfer to mammalian cells. The bacmid system of Autographa californica nuclear polyhedrosis virus (AcNPV) has already been established and widely used. However, the AcNPV does not have a potential to infect silkworm. We developed the first practical Bombyx mori nuclear polyhedrosis virus bacmid system directly applicable for the protein expression of silkworm. By using this system, the green fluorescence protein was successfully expressed in silkworm larvae and pupae not only by infection of its recombinant virus but also by direct injection of its bacmid DNA. This method provides the rapid protein production in silkworm as long as 10 days, is free from biohazard, thus will be a powerful tool for the future production factory of recombinant eukaryotic proteins and baculoviruses.
家蚕是大规模生产真核蛋白以及用于基因转移至哺乳动物细胞的重组杆状病毒最具吸引力的宿主之一。苜蓿银纹夜蛾核型多角体病毒(AcNPV)的杆粒系统已经建立并得到广泛应用。然而,AcNPV没有感染家蚕的潜力。我们开发了首个直接适用于家蚕蛋白表达的实用家蚕核型多角体病毒杆粒系统。通过使用该系统,绿色荧光蛋白不仅通过其重组病毒感染在家蚕幼虫和蛹中成功表达,还通过直接注射其杆粒DNA成功表达。这种方法可在家蚕中快速生产蛋白,只需10天,且无生物危害,因此将成为未来重组真核蛋白和杆状病毒生产工厂的有力工具。