Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system
Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system
复制标题
DOI：
10.1007/s00253-005-0127-2
复制
发表时间：
2006-06-01
影响因子：
5
通讯作者：
Maenaka, Katsumi
中科院分区：
文献类型：
--
作者：
Miao, Yungen;Zhang, Yuansong;Maenaka, Katsumi
家蚕核型多角体病毒（BmNPV）杆状病毒表达系统（BES）具有表达效率高、使用方便、饲养成本低等优点。在本报告中，我们使用了一种新开发的BmNPV杆粒，它可以同时感染B。桑细胞系和蚕幼虫。结果表明，重组杆状病毒的产生仅需7 ~ 10天，并可从小规模培养物中快速分离，然后用它来检测B。mori细胞系，相比传统的同源重组方法，其需要至少40天的时间进行多轮病毒的纯化和扩增。利用该BES，我们在BmN细胞系中表达了一个重组蜘蛛鞭毛蛋白，经SDS-PAGE和Western blot分析，其分子量约为37 kDa。家蚕杆状病毒的杆状病毒载体系统对于目的蛋白的表达具有重要的意义。
Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins.