title: High-efficiency nuclear transformation of the diatom Phaeodactylum tricornutum by electroporation.
abstract: We established a high-efficiency nuclear transformation method for the diatom Phaeodactylum tricornutum using an electroporation system. Based on a universal electroporation protocol, the conditions for the introduction of exogenous DNA including electric field strength and plasmid form were optimized. Following optimization, the diatom cells could be transformed with exogenous gene easily, the maximum transformation frequency obtained was 2.8×10(-5) cells. The cotransformation of P. tricornutum with a non-selective GUS gene together with the selectable resistance gene has also been achieved using our new method and found to be very efficient (up to 60%). The electroporation procedure described in this article offers a number of advantages, including simplicity, general utility, low-cost and high efficiency. The described method also provides some clue for developing electroporation transformation system in other eukaryotic microalgae.
authors.affiliation: Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Chunye
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China. Electronic address: hanhuahu@ihb.ac.cn.
authors.forename: Hanhua
authors.initials: H
authors.lastname: Hu
mesh_terms: D001709:Biotechnology / Q000379:methods*; D002467:Cell Nucleus / Q000235:genetics; D017377:Diatoms / Q000235:genetics*; D018274:Electroporation; D005821:Genetic Techniques; D014170:Transformation, Genetic / Q000235:genetics*
keywords: Cotransformation; Diatom transformation; Electroporation; GUS; eGFP