Transformation of Spirulina platensis strain C1 (Arthrospira sp. PCC9438) with Tn5 transposase-transposon DNA-cation liposome complex.
Spirulina platensis is one of the most commercially important species of microalgae. Thus, it is an attractive candidate for genetic manipulation and the development of novel practical applications. However, this process is hampered by the absence of a stable gene transfer system, specifically the limited number of suitable vectors and transformation methods available for this organism. Artificial transposon systems developed by extracting the essential elements from natural transposons have been extensively studied, and recently a mutated transposase and transposon system was reported to improve transformation efficiency by electroporation. We applied a modified transformation strategy using a natural Tn5 transposon, transposase, and cation liposome complex by electroporation to improve the transformation efficiency for Spirulina platensis strain C1 (Arthrospira sp. PCC9438). Aggregation of cells became visible after 3 weeks during 2.0 microg/ml chloramphenicol selection, and growth continued for more than 12 months. Transfected chloramphenicol acetyltransferase (CAT) genes were detected in the genomic DNA by Southern hybridization. Transformed cells demonstrated CAT activity, but non-transformed cells did not.
Kawata
Yoshikazu
Y
Special Division for Green Life Technology, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan, y-kawata@aist.go.jp.
Yano
Shin'ichi
S
Kojima
Hiroyuki
H
Toyomizu
Masaaki
M
D001483:Base Sequence; D015139:Blotting, Southern; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism; D000458:Cyanobacteria / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics; D018014:Gene Transfer Techniques; D008081:Liposomes; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014170:Transformation, Genetic / Q000235:genetics*; D019895:Transposases / Q000378:metabolism
D017931:DNA Primers; D004251:DNA Transposable Elements; D008081:Liposomes; C473503:Tn5 transposase; D015500:Chloramphenicol O-Acetyltransferase; D019895:Transposases