論文 - 詳細
| RRC ID | 55714 |
|---|---|
| 著者 | Xxxx P, Minamihata K, Tatsuke T, Lee JM, Kusakabe T, Kamiya N. |
| タイトル | Expression and Activation of Horseradish Peroxidase-Protein A/G Fusion Protein in Silkworm Larvae for Diagnostic Purposes. |
| ジャーナル | Biotechnol J |
| Abstract |
Recombinant protein production can create artificial proteins with desired functions by introducing genetic modifications to the target proteins. Horseradish peroxidase (HRP) has been used extensively as a reporter enzyme in biotechnological applications; however, recombinant production of HRP has not been very successful, hampering the utilization of HRP with genetic modifications. A fusion protein comprising an antibody binding protein and HRP will be an ideal bio-probe for high-quality HRP-based diagnostic systems. A HRP-protein A/G fusion protein (HRP-pAG) is designed and its production in silkworm (Bombyx mori) is evaluated for the first time. HRP-pAG is expressed in a soluble apo form, and is activated successfully by incubating with hemin. The activated HRP-pAG is used directly for ELISA experiments and retains its activity over 20 days at 4 °C. Moreover, HRP-pAG is modified with biotin by the microbial transglutaminase (MTG) reaction. The biotinylated HRP-pAG is conjugated with streptavidin to form a HRP-pAG multimer and the multimeric HRP-pAG produced higher signals in the ELISA system than monomeric HRP-pAG. The successful production of recombinant HRP in silkworm will contribute to creating novel HRP-based bioconjugates as well as further functionalization of HRP by applying enzymatic post-translational modifications. |
| 巻・号 | 13(6) |
| ページ | e1700624 |
| 公開日 | 2018-6-1 |
| DOI | 10.1002/biot.201700624 |
| PMID | 29717548 |
| MeSH | Animals Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* Biotechnology Biotin / chemistry Biotin / metabolism Biotinylation Bombyx / genetics Bombyx / metabolism* Horseradish Peroxidase / chemistry Horseradish Peroxidase / genetics Horseradish Peroxidase / metabolism* Larva / genetics Larva / metabolism Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism* Spectrophotometry, Ultraviolet Staphylococcal Protein A / chemistry Staphylococcal Protein A / genetics Staphylococcal Protein A / metabolism* Streptavidin / chemistry Streptavidin / metabolism |
| IF | 3.543 |
| 引用数 | 2 |
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オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| カイコ | f38 |