A Novel Robust Heat-inducible Promoter for Heterologous Gene Expression in Tetrahymena thermophila
文献类型：期刊论文
| 作者 | Yu, Ting1,2; Barchetta, Sabrina1; Pucciarelli, Sandra1; La Terza, Antonietta1; Miceli, Cristina1 |
| 刊名 | PROTIST
 |
| 出版日期 | 2012-03-01 |
| 卷号 | 163期号:2页码:284-295 |
| 关键词 | Cytoplasmic Hsp70 inducible promoter heterologous expression Tetrahymena thermophila |
| ISSN号 | 1434-4610 |
| 通讯作者 | Yu, T (reprint author), Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy |
| 中文摘要 | An increasing amount of data has revealed the importance of inducible promoters in ciliate research and in ciliate-related industries. However, knowledge about these promoters and related genes is relatively sparse. Here we report a novel inducible promoter from a Tetrahymena cytoplasmic Hsp70 gene member, HSP70-2. The reported promoter was able to induce the endogenous gene up to similar to 9000-fold after a short heat shock treatment and this remarkable feature has been retained when a relatively short region of the promoter was introduced into a reporter construct followed by transformation. During the recovery period following a short heat shock, both the mRNA and protein levels of the reporter gene were maintained high up to two hours. A constant heat shock treatment to the transformed cells led to a stabilization of the reporter mRNA up to at least six hours and the reporter protein continued to accumulate up to around three hours. The promoter strength appears to be similar to that of the cadmium-induced metallothionein gene ( MTT1) promoter. Therefore, the HSP70-2 promoter represents an attractive alternative for the over-expression of proteins in Tetrahymena, and the promoter-reporter gene construct used in this study is an ideal tool to help in understanding the regulation mechanisms of heat shock genes in ciliates. (C) 2011 Elsevier GmbH. All rights reserved. |
| 英文摘要 | An increasing amount of data has revealed the importance of inducible promoters in ciliate research and in ciliate-related industries. However, knowledge about these promoters and related genes is relatively sparse. Here we report a novel inducible promoter from a Tetrahymena cytoplasmic Hsp70 gene member, HSP70-2. The reported promoter was able to induce the endogenous gene up to similar to 9000-fold after a short heat shock treatment and this remarkable feature has been retained when a relatively short region of the promoter was introduced into a reporter construct followed by transformation. During the recovery period following a short heat shock, both the mRNA and protein levels of the reporter gene were maintained high up to two hours. A constant heat shock treatment to the transformed cells led to a stabilization of the reporter mRNA up to at least six hours and the reporter protein continued to accumulate up to around three hours. The promoter strength appears to be similar to that of the cadmium-induced metallothionein gene ( MTT1) promoter. Therefore, the HSP70-2 promoter represents an attractive alternative for the over-expression of proteins in Tetrahymena, and the promoter-reporter gene construct used in this study is an ideal tool to help in understanding the regulation mechanisms of heat shock genes in ciliates. (C) 2011 Elsevier GmbH. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Microbiology |
| 研究领域[WOS] | Microbiology |
| 关键词[WOS] | REPRESSIBLE PROMOTER ; PROTEIN-SYNTHESIS ; SHOCK ; HSP70 ; RNA ; PYRIFORMIS ; STRESS ; THERMOTOLERANCE ; CELLS ; PCR |
| 收录类别 | SCI |
| 资助信息 | Italian Ministry of University and Research (MIUR) |
| 语种 | 英语 |
| WOS记录号 | WOS:000299812800009 |
| 公开日期 | 2012-04-01 |
| 源URL | [http://ir.ihb.ac.cn/handle/342005/16727]  |
| 专题 | 水生生物研究所_淡水生态学研究中心_期刊论文 |
| 作者单位 | 1.Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy 2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yu, Ting,Barchetta, Sabrina,Pucciarelli, Sandra,et al. A Novel Robust Heat-inducible Promoter for Heterologous Gene Expression in Tetrahymena thermophila[J]. PROTIST,2012,163(2):284-295. |
| APA | Yu, Ting,Barchetta, Sabrina,Pucciarelli, Sandra,La Terza, Antonietta,&Miceli, Cristina.(2012).A Novel Robust Heat-inducible Promoter for Heterologous Gene Expression in Tetrahymena thermophila.PROTIST,163(2),284-295. |
| MLA | Yu, Ting,et al."A Novel Robust Heat-inducible Promoter for Heterologous Gene Expression in Tetrahymena thermophila".PROTIST 163.2(2012):284-295. |
入库方式： OAI收割
来源：水生生物研究所
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