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Development of a rapid and efficient BmNPV baculovirus expression system for application in mulberry silkworm, Bombyx mori
| Content Provider | Semantic Scholar |
|---|---|
| Author | Cao, Cuiping Wu, Xiao-Feng Zhao, Na Yao, Huipeng Lu, Xingmeng Tan, Yeping |
| Copyright Year | 2006 |
| Abstract | Silkworm-baculovirus gene expression system is one of the most powerful eukaryotic expression systems. However, due to very low recombination frequency, the traditional method to construct and obtain pure recombinant baculovirus requires plaque assay that is time-consuming and also utilizes skillful techniques. In order to overcome this disadvantage, a rapid BmNPV expression system applicable to silkworm was constructed based on the working principle of AcMNPV Bac-to-Bac system. A large 8.6 kb fragment containing the low-copy-number mini-F replicon, a kanamycin resistance marker and a segment of DNA encoding the lacZa peptide from the AcMNPV bacmid, was cloned into polyhedrin locus of BmNPV genome to replace the polyhedrin gene. This recombinant, designated as BmBacmid, was transformed into Escherichia coli DH10β strain, in which a helper plasmid encoding the transposase was already transformed. We designated the DH10β strain containing BmBacmid and helper as DH1OBmBac. With this bacterium, the recombinant baculovirus can be rapidly and easily generated through gene transposition. This system has an advantage of high recombination frequency, simple manipulation, high-efficiency and is time-saving. This approach permits large potential value in recombinant protein production using silkworm as a 'biofactory' in the future biotechnological industry and can become a powerful tool for structural and functional analysis of protein in post-genomic era. |
| Starting Page | 1692 |
| Ending Page | 1697 |
| Page Count | 6 |
| File Format | PDF HTM / HTML |
| Volume Number | 91 |
| Alternate Webpage(s) | http://www.iisc.ernet.in/currsci/dec252006/1692.pdf |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
