Establishment of a Genetic Transformation System for the Marine Pennate Diatom Fistulifera sp. Strain JPCC DA0580-A High Triglyceride Producer

Masaki Muto, Yorikane Fukuda, Michiko Nemoto, Tomoko Yoshino, Tadashi Matsunaga, Tsuyoshi Tanaka

Research output: Contribution to journalArticle

52 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)48-55
Number of pages8
JournalMarine Biotechnology
Volume15
Issue number1
DOIs
Publication statusPublished - 2013 Feb
Externally publishedYes

Fingerprint

Kanamycin Kinase
Genetic Transformation
Diatoms
Bacillariophyceae
genetic transformation
kanamycin kinase
Triglycerides
diatom
triacylglycerols
promoter regions
Microalgae
Genetic Techniques
Tungsten
Biofuels
gene
tungsten
Green Fluorescent Proteins
Genes
biodiesel
genome

Keywords

  • Biodiesel fuel
  • Diatom
  • Genetic transformation

ASJC Scopus subject areas

  • Biotechnology
  • Aquatic Science

Cite this

Establishment of a Genetic Transformation System for the Marine Pennate Diatom Fistulifera sp. Strain JPCC DA0580-A High Triglyceride Producer. / Muto, Masaki; Fukuda, Yorikane; Nemoto, Michiko; Yoshino, Tomoko; Matsunaga, Tadashi; Tanaka, Tsuyoshi.

In: Marine Biotechnology, Vol. 15, No. 1, 02.2013, p. 48-55.

Research output: Contribution to journalArticle

Muto, Masaki ; Fukuda, Yorikane ; Nemoto, Michiko ; Yoshino, Tomoko ; Matsunaga, Tadashi ; Tanaka, Tsuyoshi. / Establishment of a Genetic Transformation System for the Marine Pennate Diatom Fistulifera sp. Strain JPCC DA0580-A High Triglyceride Producer. In: Marine Biotechnology. 2013 ; Vol. 15, No. 1. pp. 48-55.
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