Reference - Detail
| RRC ID | 22783 |
|---|---|
| Author | Mon H, Lee J, Fukushima M, Nagata Y, Fujii M, Xu J, Nishi O, Iiyama K, Kusakabe T. |
| Title | Production and characterization of the celery mismatch endonuclease CEL II using baculovirus/silkworm expression system. |
| Journal | 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. |
| Volume | 97(15) |
| Pages | 6813-22 |
| Published | 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 |
| Times Cited | 10 |
| WOS Category | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 4 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Silkworms | d17 |