RRC ID 3449
Author Miao Y, Zhang Y, Nakagaki K, Zhao T, Zhao A, Meng Y, Nakagaki M, Park EY, Maenaka K.
Title Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system.
Journal Appl. Microbiol. Biotechnol.
Abstract 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.
Volume 71(2)
Pages 192-9
Published 2006-6
DOI 10.1007/s00253-005-0127-2
PMID 16158284
MeSH Amino Acid Sequence Animals Baculoviridae / genetics* Base Sequence Biotechnology / methods* Blotting, Western Bombyx / metabolism Bombyx / virology* Cloning, Molecular DNA / chemistry DNA / genetics Molecular Sequence Data Polymerase Chain Reaction Proteins / genetics Proteins / metabolism* Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Spiders / genetics* Spiders / metabolism
IF 3.34
Times Cited 55
Silkworms ?