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Development of an in vivo fluorescence based gene expression reporter system for Clostridium tyrobutyricum
- Source :
- Journal of Biotechnology. 305:18-22
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- C. tyrobutyricum, an acidogenic Clostridium, has aroused increasing interest due to its potential to produce biofuel efficiently. However, construction of recombinant C. tyrobutyricum for enhanced biofuel production has been impeded by the limited genetic engineering tools. In this study, a flavin mononucleotide (FMN)-dependent fluorescent protein Bs2-based gene expression reporter system was developed to monitor transformation and explore in vivo strength and regulation of various promoters in C. tyrobutyricum and C. acetobutylicum. Unlike green fluorescent protein (GFP) and its variants, Bs2 can emit green light without oxygen, which makes it extremely suitable for promoter screening and transformation confirmation in organisms grown anaerobically. The expression levels of bs2 under thiolase promoters from C. tyrobutyricum and C. acetobutylicum were measured and compared based on fluorescence intensities. The capacities of the two promoters in driving secondary alcohol dehydrogenase (adh) gene for isopropanol production in C. tyrobutyricum were distinguished, confirming that this reporter system is a convenient, effective and reliable tool for promoter strength assay and real time monitoring in C. tyrobutyricum, while demonstrating the feasibility of producing isopropanol in C. tyrobutyricum for the first time.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Flavin Mononucleotide
Flavin mononucleotide
Bioengineering
01 natural sciences
Applied Microbiology and Biotechnology
Fluorescence
Green fluorescent protein
2-Propanol
03 medical and health sciences
chemistry.chemical_compound
Clostridium
Genes, Reporter
010608 biotechnology
Gene expression
Promoter Regions, Genetic
biology
Thiolase
Alcohol Dehydrogenase
Promoter
General Medicine
biology.organism_classification
Clostridium tyrobutyricum
Transformation (genetics)
030104 developmental biology
chemistry
Biochemistry
Biofuels
Clostridium acetobutylicum
Genetic Engineering
Biotechnology
Subjects
Details
- ISSN :
- 01681656
- Volume :
- 305
- Database :
- OpenAIRE
- Journal :
- Journal of Biotechnology
- Accession number :
- edsair.doi.dedup.....ac3ac0648a3f2467c3b688f1deb6878a