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A Novel Robust Heat-inducible Promoter for Heterologous Gene Expression in Tetrahymena thermophila
Yu, Ting1,2; Barchetta, Sabrina1; Pucciarelli, Sandra1; La Terza, Antonietta1; Miceli, Cristina1; Yu, T (reprint author), Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy
2012-03-01
Source PublicationPROTIST
ISSN1434-4610
Volume163Issue:2Pages:284-295
AbstractAn 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.; 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.
SubtypeArticle
KeywordCytoplasmic Hsp70 Inducible Promoter Heterologous Expression Tetrahymena Thermophila
Department[Yu, Ting; Barchetta, Sabrina; Pucciarelli, Sandra; La Terza, Antonietta; Miceli, Cristina] Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy; [Yu, Ting] Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China
DOI10.1016/j.protis.2011.07.003
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationItalian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR)
Indexed BySCI
Language英语
WOS Research AreaMicrobiology
WOS SubjectMicrobiology
WOS IDWOS:000299812800009
WOS KeywordREPRESSIBLE PROMOTER ; PROTEIN-SYNTHESIS ; SHOCK ; HSP70 ; RNA ; PYRIFORMIS ; STRESS ; THERMOTOLERANCE ; CELLS ; PCR
Funding OrganizationItalian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR) ; Italian Ministry of University and Research (MIUR)
Citation statistics
Cited Times:10[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/16727
Collection淡水生态学研究中心_期刊论文
Corresponding AuthorYu, T (reprint author), Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy
Affiliation1.Univ Camerino, Sch Biosci & Biotechnol, I-62032 Camerino, Italy
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China
Recommended Citation
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,&Yu, T .(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.
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