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Expression of a synthetic gene encoding canine milk lysozyme in Escherichia coli and characterization of the expressed protein
https://ir.soken.ac.jp/records/4289
https://ir.soken.ac.jp/records/428993ad5503-de40-42af-a780-7389a5be5693
| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||
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| 公開日 | 2014-03-12 | |||||||||||
| タイトル | ||||||||||||
| タイトル | Expression of a synthetic gene encoding canine milk lysozyme in Escherichia coli and characterization of the expressed protein | |||||||||||
| タイトル | ||||||||||||
| タイトル | Expression of a synthetic gene encoding canine milk lysozyme in Escherichia coli and characterization of the expressed protein | |||||||||||
| 言語 | en | |||||||||||
| 言語 | ||||||||||||
| 言語 | eng | |||||||||||
| 資源タイプ | ||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||
| 資源タイプ | journal article | |||||||||||
| アクセス権 | ||||||||||||
| アクセス権 | metadata only access | |||||||||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||||||||
| 著者 |
KOSHIBA, Takumi
× KOSHIBA, Takumi
× KUWAJIMA, Kunihiro
× et, al.
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| 著者別名 |
桑島, 邦博
× 桑島, 邦博
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| 抄録 | ||||||||||||
| 内容記述タイプ | Abstract | |||||||||||
| 内容記述 | A high-expression plasmid of the canine milk lysozyme, which belongs to the family of calcium-binding lysozymes, was constructed in order to study its physico-chemical properties. Because the cDNA sequence of the protein has not yet been determined, a 400 base-pair gene encoding canine milk lysozyme was first designed on the basis of the known amino acid sequence. The gene was constructed by an enzymatic assembly of 21 chemically synthesized oligonucleotides and inserted into an Escherichia coli expression vector by stepwise ligation. The expression plasmid thus constructed was transformed into BL21(DE3)/pLysS cells. The gene product accumulated as inclusion bodies in an insoluble fraction. Recombinant canine milk lysozyme was obtained by purification and refolding of the product and showed the same characteristics in terms of bacteriolytic activity and far- and near-UV circular dichroism spectra as the authentic protein. The NMR spectra of refolded lysozyme were also characteristic of a native globular protein. It was concluded that recombinant canine milk lysozyme was folded into the correct native structure. Moreover, the thermal unfolding profiles of the refolded recombinant lysozyme showed a stable equilibrium intermediate, indicating that the molten globule state of this protein was extraordinarily stable. This expression system of canine milk lysozyme will enable biophysical and structural studies of this protein to be extended. | |||||||||||
| 書誌情報 |
Protein Engineering, Design and Selection en : Protein Engineering, Design and Selection 巻 12, 号 5, p. 429-435, 発行日 1999 |
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| 出版者 | ||||||||||||
| 出版者 | Oxford University Press | |||||||||||
| ISSN | ||||||||||||
| 収録物識別子タイプ | ISSN | |||||||||||
| 収録物識別子 | 1741-0126 | |||||||||||
| DOI | ||||||||||||
| 識別子タイプ | DOI | |||||||||||
| 関連識別子 | https://doi.org/10.1093/protein/12.5.429 | |||||||||||
| 関連名称 | 10.1093/protein/12.5.429 | |||||||||||
| 権利 | ||||||||||||
| 権利情報 | © Oxford University Press | |||||||||||
