論文 - 詳細
| RRC ID | 506 |
|---|---|
| 著者 | He N, Fujii H, Kusakabe T, Aso Y, Banno Y, Yamamoto K. |
| タイトル | Overexpression in Escherichia coli and purification of recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm. |
| ジャーナル | Protein Expr Purif |
| Abstract |
Present research provided an efficient approach to obtain large quantities of active recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm, Bombyx mori. The cDNA encoding mature CI-b1 was cloned into pDEST17 vector. Recombinant protein with hexa-histidine tag attached to the N-terminal of CI-b1 was expressed in Escherichia coli Origami B cells. It can be purified to homogeneity via the gel filtration chromatography on a Sephacryl S-200 column followed the affinity chromatography on a Ni-NTA column. The two sequential purification procedures yielded 4.3mg purified (His)(6)-tagged CI-b1 from 200ml of culture medium. Studies on (His)(6)-tagged CI-b1 revealed that three disulfide bonds were formed in the recombinant CI-b1 and the inhibitory properties of recombinant CI-b1 against alpha-chymotrypsin were similar to those of native CI-b1. Recombinant CI-b1 immobilized on Ni-NTA resin was used to detect the interactions occurring between the CI-b1 and its target factors. |
| 巻・号 | 38(1) |
| ページ | 9-16 |
| 公開日 | 2004-11-1 |
| DOI | 10.1016/j.pep.2004.07.010 |
| PII | S1046-5928(04)00260-8 |
| PMID | 15477076 |
| MeSH | Animals Bombyx / metabolism* Chromatography, Affinity Chymotrypsin / antagonists & inhibitors Escherichia coli / genetics* Escherichia coli / metabolism Molecular Sequence Data Molecular Weight Peptides / genetics* Plant Proteins / genetics* Recombinant Proteins / genetics Recombinant Proteins / isolation & purification* Recombinant Proteins / metabolism |
| IF | 1.513 |
| 引用数 | 4 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| カイコ | p50 |