Construction of a cysteine protease deficient Bombyx mori multiple nucleopolyhedrovirus bacmid and its application to improve expression of a fusion protein
Construction of a cysteine protease deficient Bombyx mori multiple nucleopolyhedrovirus bacmid and its application to improve expression of a fusion protein
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DOI：
10.1016/j.jviromet.2007.04.005
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发表时间：
2007-09-01
影响因子：
3.1
通讯作者：
Park, Enoch Y.
中科院分区：
文献类型：
--
作者：
Hiyoshi, Masato;Kageshima, Ayano;Park, Enoch Y.
利用λ重组系统构建了BmMNPV半胱氨酸蛋白酶基因缺失的杆状病毒（CPD BmMNPV bacmid）。用该CPD-BmMNPV杆粒感染家蚕细胞和家蚕幼虫，其蛋白酶活性分别降低了94%和85%。通过使用该系统，GFP（uv）-β 1，3-N-乙酰氨基葡萄糖转移酶2（GFP（uv）-β 3GnT 2）融合蛋白在家蚕幼虫中成功表达，蛋白质降解较少且不发生幼虫液化，β 3GnT活性提高了30%。该CPD-BmMNPV杆粒系统提供了在蚕中的快速蛋白质生产，并且可用于生产重组真核蛋白而不发生蛋白水解降解。(c)2007 Elsevier B. V.保留所有权利。
The bacmid system of BmMNPV with cysteine protease gene deletion (CPD-BmMNPV bacmid) was constructed using the lambda recombination system. The protease activities of Bombyx mori cells and silkworm larvae infected with this CPD-BmMNPV bacmid were reduced by 94% and 85%, respectively. By using this system, a GFP(uv)-beta 1,3-N-acetylglucosaminyltransferase 2 (GFP(uv)-beta 3GnT2) fusion protein was successfully expressed in silkworm larvae with less protein degradation and without larvae liquefaction; beta 3GnT activity improved 30%. This CPD-BmMNPV bacmid system provides rapid protein production in silkworms and can be used for the production of recombinant eukaryotic proteins without proteolytic degradation. (c) 2007 Elsevier B.V. All rights reserved.