부산대학교 도서관

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 lacZ a 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 DH10 b strain, in which a helper plasmid encoding the transposase was already transformed. We designated the DH10 b strain containing BmBacmid and helper as DH10BmBac. 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. [ABSTRACT FROM AUTHOR]