Overcoming Intrinsic Restriction Enzyme Barriers Enhances Transformation Efficiency in Arthrospira platensis C1.
The development of a reliable genetic transformation system for Arthrospira platensis has been a long-term goal, mainly for those trying either to improve its performance in large-scale cultivation systems or to enhance its value as food and feed additives. However, so far, most of the attempts to develop such a transformation system have had limited success. In this study, an efficient and stable transformation system for A. platensis C1 was successfully developed. Based on electroporation and transposon techniques, exogenous DNA could be transferred to and stably maintained in the A. platensis C1 genome. Most strains of Arthrospira possess strong restriction barriers, hampering the development of a gene transfer system for this group of cyanobacteria. By using a type I restriction inhibitor and liposomes to protect the DNA from nuclease digestion, the transformation efficiency was significantly improved. The transformants were able to grow on a selective medium for more than eight passages, and the transformed DNA could be detected from the stable transformants. We propose that the intrinsic endonuclease enzymes, particularly the type I restriction enzyme, in A. platensis C1 play an important role in the transformation efficiency of this industrial important cyanobacterium.
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi (Bang Khun Thian), Bangkok, Thailand.
Wattana
W
Jeamton
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi (Bang Khun Thian), Bangkok, Thailand.
Sudarat
S
Dulsawat
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi (Bang Khun Thian), Bangkok, Thailand.
Morakot
M
Tanticharoen
The Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, Sede Boqer campus, Israel.
Avigad
A
Vonshak
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi (Bang Khun Thian), Bangkok, Thailand.|School of Bioresources and Technology, King Mongkut's University of Technology Thonburi (Bang Khun Thian), Bangkok, Thailand.
Supapon
S
Cheevadhanarak
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004251:DNA Transposable Elements; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D016680:Genome, Bacterial; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015203:Reproducibility of Results; D000198:Spectinomycin / Q000494:pharmacology; D053545:Spirulina / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D014170:Transformation, Genetic; D019895:Transposases / Q000235:genetics
D001426:Bacterial Proteins; D003470:Culture Media; D004251:DNA Transposable Elements; D004798:Enzymes; C473503:Tn5 transposase; D000198:Spectinomycin; D019895:Transposases
Arthrospira platensis; Cyanobacteria; Liposome; Nuclease; Transformation efficiency