Flavin mononucleotide (FMN)-based fluorescent protein (FbFP) as reporter for promoter screening in Clostridium cellulolyticum
Teng, Lin1,2; Wang, Kun3; Xu, Jian1; Xu, Chenggang1
刊名JOURNAL OF MICROBIOLOGICAL METHODS
2015-12-01
卷号119期号:1页码:37-43
关键词Clostridium cellulolyticum FbFP Promoter screening Constitutive Inducible
英文摘要

Conventional methods for screening promoters in anaerobic bacteria are generally based on detection of enzymatic reactions and thus usually complicated or strain specific. Therefore a more efficient and universal method will be valuable. Here, using cellulolytic bacteria Clostridium cellulolyticum H10 as a model, we employed an oxygen-independent flavin-based fluorescent protein (FbFP) derived from Pseudomonas putida as a quantitative reporter for the screening of promoter via monitoring fluorescence intensity. The stability and reliability of FbFP fluorescence were proven by the high correlation (R-2 = 0.87) between fluorescence intensity and abundance of FbFP. Moreover, two endogenous promoters with exceptional performance were identified and characterized, including a constitutive promoter p3398 and an inducible promoter p1133. Compared to the existing reporter systems widely used in clostridia, this FbFP-based method is more rapid, intuitive and versatile, and the endogenous promoters reported here should enrich the synthetic biology toolbox for this and related organisms. (C) 2015 Elsevier B.V. All rights reserved.

WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods ; Microbiology
研究领域[WOS]Biochemistry & Molecular Biology ; Microbiology
关键词[WOS]GENE-EXPRESSION SYSTEM ; ACETOBUTYLICUM ; GFP ; PERFRINGENS ; IMPROVEMENT ; DISRUPTION ; BIOMASS ; MODEL ; H10
收录类别SCI
语种英语
WOS记录号WOS:000366442400007
内容类型期刊论文
源URL[http://ir.qibebt.ac.cn/handle/337004/6840]  
专题青岛生物能源与过程研究所_单细胞中心
作者单位1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Single Cell Ctr, CAS Key Lab Biofuels,Shandong Key Lab Energy Gene, Qingdao 266101, Shandong, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Southwest Univ, Coll Hort & Landscape Architecture, Minist Educ, Key Lab Hort Sci Southern Mountainous Reg, Chongqing 400716, Peoples R China
推荐引用方式
GB/T 7714
Teng, Lin,Wang, Kun,Xu, Jian,et al. Flavin mononucleotide (FMN)-based fluorescent protein (FbFP) as reporter for promoter screening in Clostridium cellulolyticum[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2015,119(1):37-43.
APA Teng, Lin,Wang, Kun,Xu, Jian,&Xu, Chenggang.(2015).Flavin mononucleotide (FMN)-based fluorescent protein (FbFP) as reporter for promoter screening in Clostridium cellulolyticum.JOURNAL OF MICROBIOLOGICAL METHODS,119(1),37-43.
MLA Teng, Lin,et al."Flavin mononucleotide (FMN)-based fluorescent protein (FbFP) as reporter for promoter screening in Clostridium cellulolyticum".JOURNAL OF MICROBIOLOGICAL METHODS 119.1(2015):37-43.
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