Fine-Tuning of Photoautotrophic Protein Production by Combining Promoters and Neutral Sites in the Cyanobacterium Synechocystis sp. Strain PCC 6803
Autor: | Himadri B. Pakrasi, Andrew H. Ng, Bertram M. Berla |
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Rok vydání: | 2015 |
Předmět: |
Cyanobacteria
Light Saccharomyces cerevisiae Molecular Sequence Data Genetics and Molecular Biology Applied Microbiology and Biotechnology Metabolic engineering Plasmid Shuttle vector Bacterial Proteins Promoter Regions Genetic Gene Genetics Autotrophic Processes Ecology biology Base Sequence Synechocystis Gene Expression Regulation Bacterial biology.organism_classification Biochemistry Expression cassette Food Science Biotechnology Plasmids |
Zdroj: | Applied and environmental microbiology. 81(19) |
ISSN: | 1098-5336 |
Popis: | Cyanobacteria are photosynthetic cell factories that use solar energy to convert CO 2 into useful products. Despite this attractive feature, the development of tools for engineering cyanobacterial chassis has lagged behind that for heterotrophs such as Escherichia coli or Saccharomyces cerevisiae . Heterologous genes in cyanobacteria are often integrated at presumptively “neutral” chromosomal sites, with unknown effects. We used transcriptome sequencing (RNA-seq) data for the model cyanobacterium Synechocystis sp. strain PCC 6803 to identify neutral sites from which no transcripts are expressed. We characterized the two largest such sites on the chromosome, a site on an endogenous plasmid, and a shuttle vector by integrating an enhanced yellow fluorescent protein (EYFP) expression cassette expressed from either the P cpc560 or the P trc1O promoter into each locus. Expression from the endogenous plasmid was as much as 14-fold higher than that from the chromosome, with intermediate expression from the shuttle vector. The expression characteristics of each locus correlated predictably with the promoters used. These findings provide novel, characterized tools for synthetic biology and metabolic engineering in cyanobacteria. |
Databáze: | OpenAIRE |
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