Park Enoch Y, Abe Takahiro, Kato Tatsuya
Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Biotechnol Appl Biochem. 2008 Feb;49(Pt 2):135-40. doi: 10.1042/BA20070098.
A bacmid system of BmMNPV [Bombyx mori (silkworm) multiple nucleopolyhedrovirus] with both the cysteine-protease and chitinase genes deleted (BmMNPV-CP(-)-Chi(-)) was constructed. The viral protease and chitinase activities in the haemolymph of Bombyx mori larvae infected with this BmMNPV-CP(-)-Chi(-) bacmid were reduced by 95 and 50% respectively. By using this system, a GFP(uv)-beta3GnT2 (green fluorescent protein excited by UV light-beta1,3-N-acetylglucosaminyltransferase 2) fusion protein was successfully expressed in silkworm larvae with less protein degradation and without larvae liquefaction; beta3GnT2 activity improved 2.8-fold over that of unmodified bacmid. This BmMNPV-CP(-)-Chi(-) bacmid system provides rapid protein production in silkworms and can be used for the production of recombinant eukaryotic proteins without proteolytic degradation.
构建了一种缺失半胱氨酸蛋白酶和几丁质酶基因的家蚕多核型多角体病毒（BmMNPV）杆粒系统（BmMNPV-CP(-)-Chi(-)）。感染这种BmMNPV-CP(-)-Chi(-)杆粒的家蚕幼虫血淋巴中的病毒蛋白酶和几丁质酶活性分别降低了95%和50%。利用该系统，绿色荧光蛋白（UV激发型）-β3GnT2（β1,3-N-乙酰葡糖胺基转移酶2）融合蛋白在蚕幼虫中成功表达，蛋白质降解较少且无幼虫液化现象；β3GnT2活性比未修饰的杆粒提高了2.8倍。这种BmMNPV-CP(-)-Chi(-)杆粒系统可在家蚕中快速生产蛋白质，可用于生产无蛋白水解降解的重组真核蛋白。
