RRC ID 22783
著者 Mon H, Lee J, Fukushima M, Nagata Y, Fujii M, Xu J, Nishi O, Iiyama K, Kusakabe T.
タイトル Production and characterization of the celery mismatch endonuclease CEL II using baculovirus/silkworm expression system.
ジャーナル Appl Microbiol Biotechnol
Abstract Mutation and polymorphism detection by nucleases has become a more important tool in clinical and biological researches. There are several kinds of single-stranded nucleases for detecting mismatched DNAs. One of them, CEL II, was isolated from Apium graveolens and cleaves DNA with high specificity at sites of mismatch. High-throughput mutation scanning requires large quantity of CEL II endonuclease. Here, we demonstrate high-level expression of CEL II using silkworm-baculovirus system. The recombinant CEL II secreted in silkworm hemolymph was glycosylated and susceptible to N-glycosidase F. Additionally, larger metal ions such as Ca(2+) and Sr(2+) were able to replace Mg(2+) and enhanced mismatch cleavage activity of CEL II. These results indicate that the silkworm-baculovirus platform is a good alternative system to obtain the functional CEL II.
巻・号 97(15)
ページ 6813-22
公開日 2013-8-1
DOI 10.1007/s00253-012-4583-1
PMID 23179626
MeSH Animals Apium / enzymology* Baculoviridae / genetics* Base Sequence Bombyx / genetics* DNA Primers DNA Restriction Enzymes / metabolism* Glycosylation
IF 3.53
引用数 10
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
カイコ d17