Production and characterization of the celery mismatch endonuclease CEL II using baculovirus/silkworm expression system

Hiroaki Mon, Jaeman Lee, Mai Fukushima, Yudai Nagata, Mie Fujii, Jian Xu, Oumi Nishi, Kazuhiro Iiyama, Takahiro Kusakabe

研究成果: Contribution to journalArticle査読

11 被引用数 (Scopus)

抄録

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 Ca2+ and Sr2+ were able to replace Mg2+ 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.

本文言語英語
ページ(範囲)6813-6822
ページ数10
ジャーナルApplied Microbiology and Biotechnology
97
15
DOI
出版ステータス出版済み - 8 2013

All Science Journal Classification (ASJC) codes

  • バイオテクノロジー
  • 応用微生物学とバイオテクノロジー

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