論文 - 詳細
| RRC ID | 3801 |
|---|---|
| 著者 | Kondo N, Ebihara A, Ru H, Kuramitsu S, Iwamoto A, Rao Z, Matsuda Z. |
| タイトル | Thermus thermophilus-derived protein tags that aid in preparation of insoluble viral proteins. |
| ジャーナル | Anal Biochem |
| Abstract |
The expression and solubilization of insoluble proteins have been facilitated by the introduction of protein tags. In our analyses of viral protein R (Vpr) of human immunodeficiency virus 1 (HIV-1), however, several conventional tag proteins enhanced its expression but failed to solubilize it. Therefore, we decided to explore whether proteins derived from Thermus thermophilus HB8 (T. th.), a highly heat-stable bacterium, could be used as tag proteins to enhance the solubilization of Vpr. Based on the data accumulated during the recent structural genomics project of T. th., we selected 15 T. th. proteins with high expression levels and solubilities. From this group, we identified a T. th. tag protein that expressed Vpr in a soluble form. Furthermore, two T. th. tag proteins, including the identified one, were found to solubilize the extremely insoluble membrane-spanning domain of the envelope protein of HIV-1. When green fluorescent protein (GFP) was used as a passenger protein of T. th. tags, the brightness and stability of GFP were similar to those of untagged GFP, suggesting that the T. th. tags do not negatively affect the function of the passenger protein. Thus, data of structural genomics can be applied to generate a customized versatile protein tag for protein analyses. |
| 巻・号 | 385(2) |
| ページ | 278-85 |
| 公開日 | 2009-2-15 |
| DOI | 10.1016/j.ab.2008.10.050 |
| PII | S0003-2697(08)00771-9 |
| PMID | 19084492 |
| MeSH | Genomics / methods Green Fluorescent Proteins HIV-1 / chemistry Human Immunodeficiency Virus Proteins / analysis Humans Molecular Probe Techniques Thermus thermophilus / chemistry* Viral Proteins / analysis* |
| IF | 2.877 |
| 引用数 | 3 |
| WOS 分野 | CHEMISTRY, ANALYTICAL BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 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 | |
| 遺伝子材料 | Thermus thermophilus expression plasmids TEx12F01 (THR004921) TEx18D04 (THR007276) |