論文 - 詳細
| RRC ID | 73956 |
|---|---|
| 著者 | Inouye S. |
| タイトル | Cloning, expression, purification and characterization of an isotype of clytin, a calcium-binding photoprotein from the luminous hydromedusa Clytia gregarium. |
| ジャーナル | J Biochem |
| Abstract |
The cDNA for an isotype of clytin, a calcium-binding photoprotein from the luminous jellyfish Clytia gregarium, was identified and named clytin-II. The histidine-tagged apoprotein of clytin-II expressed into the periplasmic space of Escherichia coli cells was isolated by nickel chelate affinity chromatography. Recombinant clytin-II regenerated from apoprotein by incubation with coelenterazine was purified. The yield of purified clytin-II was 13 mg from 2 l of cultured cells with purity >95%. The luminescence properties of clytin-II were characterized by comparison with clytin-I and aequorin, and semi-synthetic clytin-II was also prepared. The initial luminescence intensity of clytin-II triggered by Ca(2+) was 4.5 times higher than that of clytin-I and aequorin, but the luminescence capacity was close to clytin-I and aequorin. Thus, clytin-II is a useful protein, showing high sensitivity in the signal-to-noise ratio of luminescence intensity. |
| 巻・号 | 143(5) |
| ページ | 711-7 |
| 公開日 | 2008-5-1 |
| DOI | 10.1093/jb/mvn024 |
| PII | mvn024 |
| PMID | 18296715 |
| MeSH | Aequorin / immunology Amino Acid Sequence Animals Antibodies Apoproteins / immunology Calcium / metabolism Calcium-Binding Proteins / chemistry* Calcium-Binding Proteins / genetics* Calcium-Binding Proteins / metabolism Cloning, Molecular Escherichia coli / genetics Hydrozoa / chemistry Hydrozoa / genetics* Immunoblotting Luminescent Proteins / chemistry* Luminescent Proteins / genetics* Luminescent Proteins / metabolism Molecular Sequence Data Recombinant Proteins / chemistry Recombinant Proteins / immunology Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Homology, Amino Acid |
| IF | 2.476 |
| リソース情報 | |
| 遺伝子材料 | pCL31 (RDB19838) |