title: Establishment of a genetic transformation system for the marine pennate diatom Fistulifera sp. strain JPCC DA0580--a high triglyceride producer.
abstract: A genetic transformation system for the marine pennate diatom, Fistulifera sp. JPCC DA0580, was established using microparticle bombardment methods. Strain JPCC DA0580 has been recently identified as the highest triglyceride (60 % w/w) producer from a culture collection of 1,393 strains of marine microalgae, and it is expected to be a feasible source of biodiesel fuel. The transformation conditions for strain JPCC DA0580 were optimised using the green fluorescent protein gene (gfp) and the gene encoding neomycin phosphotransferase II (nptII). The most efficient rate of transformation was attained when tungsten particles (0.6 μm in diameter) were used for microparticle bombardment. The effect of endogenous and exogenous promoters on the expression of nptII was examined. Endogenous promoters were more efficient for obtaining transformants compared with exogenous promoters. Southern hybridisation analysis suggested that nptII integrated into the nuclear genome. This genetic manipulation technique should allow us to understand the mechanisms of high triglyceride accumulation in this strain, thereby contributing to improving BDF production.
authors.affiliation: Graduate School of Bio-Applications and Systems Engineering, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Muto
authors.forename: Yorikane
authors.initials: Y
authors.lastname: Fukuda
authors.forename: Michiko
authors.initials: M
authors.lastname: Nemoto
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yoshino
authors.forename: Tadashi
authors.initials: T
authors.lastname: Matsunaga
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D056804:Biofuels; D019470:Biolistics / Q000379:methods; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D017377:Diatoms / Q000235:genetics* / Q000378:metabolism; D019868:Kanamycin Kinase / Q000235:genetics; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics*; D014280:Triglycerides / Q000096:biosynthesis*
chemical_list: D056804:Biofuels; D017931:DNA Primers; D014280:Triglycerides; D019868:Kanamycin Kinase