Overcoming Intrinsic Restriction Enzyme Barriers Enhances Transformation Efficiency in Arthrospira platensis C1

Jeamton, Wattana; Dulsawat, Sudarat; Tanticharoen, Morakot; Vonshak, Avigad; Cheevadhanarak, Supapon

doi:10.1093/pcp/pcx016

Cited by 28 publications

(20 citation statements)

References 49 publications

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“…However, this protocol was not reproduced. Recent study of Jeamton et al ( 2017 ), resulted in an efficient and stable transformation of Arthrospira platenis C1. This protocol was also based on electroporation, but its novelty was the use of a type I restriction-modification systems inhibitor and liposomes to protect the DNA.…”

Section: Geneticsmentioning

confidence: 99%

Edible Cyanobacterial Genus Arthrospira: Actual State of the Art in Cultivation Methods, Genetics, and Application in Medicine

et al. 2017

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The cyanobacterial genus Arthrospira appears very conserved and has been divided into five main genetic clusters on the basis of molecular taxonomy markers. Genetic studies of seven Arthrospira strains, including genome sequencing, have enabled a better understanding of those photosynthetic prokaryotes. Even though genetic manipulations have not yet been performed with success, many genomic and proteomic features such as stress adaptation, nitrogen fixation, or biofuel production have been characterized. Many of above-mentioned studies aimed to optimize the cultivation conditions. Factors like the light intensity and quality, the nitrogen source, or different modes of growth (auto-, hetero-, or mixotrophic) have been studied in detail. The scaling-up of the biomass production using photobioreactors, either closed or open, was also investigated to increase the production of useful compounds. The richness of nutrients contained in the genus Arthrospira can be used for promising applications in the biomedical domain. Ingredients such as the calcium spirulan, immulina, C-phycocyanin, and γ-linolenic acid (GLA) show a strong biological activity. Recently, its use in the fight against cancer cells was documented in many publications. The health-promoting action of “Spirulina” has been demonstrated in the case of cardiovascular diseases and age-related conditions. Some compounds also have potent immunomodulatory properties, promoting the growth of beneficial gut microflora, acting as antimicrobial and antiviral. Products derived from Arthrospira were shown to successfully replace biomaterial scaffolds in regenerative medicine. Supplementation with the cyanobacterium also improves the health of livestock and quality of the products of animal origin. They were also used in cosmetic preparations.

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Section: Geneticsmentioning

confidence: 99%

Edible Cyanobacterial Genus Arthrospira: Actual State of the Art in Cultivation Methods, Genetics, and Application in Medicine

et al. 2017

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“…Gene transfer into Nostoc sp. PCC 7120 by conjugation or into Arthrospira platensis C1 by electroporation was improved if DNA was protected from restriction endonucleases by methylation or by the use of suitable inhibitors [58,59]. Genes for single strand exonuclease RecJ [60], for several endonucleases, and for CRISPR/CAS [61][62][63] are present in the genome of Phormidium lacuna and widely distributed among other cyanobacteria including also other naturally competent species.…”

Section: Discussionmentioning

confidence: 99%

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Nies¹,

Mielke²,

Pochert³

et al. 2020

PLoS ONE

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Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. Applications are most often based on recombinant work, and the establishment of transformation can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the genome by homologous recombination. This is the first example for natural transformation filamentous non-heterocystous cyanobacterium. We found that Phormidium lacuna is polyploid, each cell has about 20-90 chromosomes. Transformed filaments were resistant against up to 14 mg/ml of kanamycin. Formerly, natural transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular species. Our finding suggests that natural competence is more distributed among cyanobacteria than previously thought. This is supported by bioinformatic analyses which show that all protein factors for natural transformation are present in the majority of the analyzed cyanobacteria.

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“…Homogenization of filaments and subsequent washings could remove extracellular polymeric materials and nucleases (5) and could thereby promote NT. Members of the genus Arthrospira (or Spirulina ), for which high nuclease activities were reported (40, 51), might be transformed by the same or a similar approach. Trials with other cyanobacteria that follow either the standard protocol or a simplified version of it are easy and can rapidly provide clear results.…”

Section: Discussionmentioning

confidence: 99%

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Nies

Mielke

Pochert

et al. 2019

Preprint

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9Research for biotechnological applications of cyanobacteria focuses on synthetic pathways and 10 bioreactor design, while little effort is devoted to introduce new, promising organisms in the field. 11Applications are most often based on recombinant work, and the establishment of transformation 12 can be a risky, time-consuming procedure. In this work we demonstrate the natural transformation 13 of the filamentous cyanobacterium Phormidium lacuna and insertion of a selection marker into the 14 genome by homologous integration. This is the first example for natural transformation of a member 15 of the order Oscillatoriales. We found that Phormidium lacuna is polyploid, each cell has about 20-16 100 chromosomes. Transformed filaments were resistant against up to 15 mg/ml of kanamycin, and 17 the high resistance feature allowed for rapid segregation into all chromosomes. Formerly, natural 18 transformation in cyanobacteria has been considered a rare and exclusive feature of a few unicellular 19 species. Our finding suggests that natural competence is more distributed among cyanobacteria than 20 previously thought. This is supported by bioinformatic analyses which show that all protein factors 21 for natural transformation are present in the majority of the analyzed cyanobacteria. 22

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Overcoming Intrinsic Restriction Enzyme Barriers Enhances Transformation Efficiency in Arthrospira platensis C1

Cited by 28 publications

References 49 publications

Edible Cyanobacterial Genus Arthrospira: Actual State of the Art in Cultivation Methods, Genetics, and Application in Medicine

Edible Cyanobacterial Genus Arthrospira: Actual State of the Art in Cultivation Methods, Genetics, and Application in Medicine

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

Natural transformation of the filamentous cyanobacterium Phormidium lacuna

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