Highly Efficient Transformation of the DiatomPhaeodactylum tricornutumby Multi-Pulse Electroporation

High-efficiency nuclear transformation of the diatom Phaeodactylum tricornutum by electroporation

“…1B). A large enhancement of the transformation frequency by the addition of carrier DNA and using a linearized vector has been reported in microalgae (Kilian et al, 2011;Miyahara et al, 2013;Shimogawara et al, 1998;Radakovits et al, 2012). The maximal transformation efficiency reached about 2.8 × 10…”

“…However, a detailed protocol was not presented in their paper. Miyahara et al (2013) also reported electroporation transformation of P. tricornutum, however, the protocol applied a multi-pulse electroporation for the transformation, and the efficiency depends on the number of poring pulses applied. Although the transformation efficiency is a little bit higher than the achieved efficiency by our protocol, the multi-pulse electroporation are dependent on a special apparatus and it is not routinely available in most laboratories.…”

“…(Kilian et al, 2011;Radakovits et al, 2012) and Scenedesmus obliquus (Guo et al, 2013). Niu et al (2012) and Miyahara et al (2013) reported the transformation of P. tricornutum by electroporation, however, the former did not achieve higher transformation efficiency compared with microprojectile bombardment while the latter applied a multi-pulse electroporation system dependent on a special apparatus. In this study, we developed a simple method for high efficiency nuclear transformation of P. tricornutum by electroporation.…”

Development of a new constitutive expression system for the transformation of the diatom Phaeodactylum tricornutum

“…M17 (1.1 μm in diameter) tungsten particles were coated with plasmids linearized by PsiI and ScaI digestion [15,16]. Particle bombardment of P. tricornutum cells was performed using a Biolistic Particle Delivery System PDS-1000/He (Bio-Rad Laboratories, USA) fitted with 1550 psi rupture discs as recommended by the manufacturer.…”

High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product

“…In addition, transformation frequencies in the two N. oceanica strains were 16.3-32 × 10 −6 , which are far higher than that in the same species strain W2J3B (2.5 × 10 −6 ) reported by Kilian et al 16) Recently, highly efficient transformation by electroporation has been established in the heterokont, Phaeodactylum tricornutum. 35) This transformation system used a linearized vector to achieve a high transformation efficiency of 45 × 10 −6 by a multi-pulse electroporation. In plant cells, transformation using PCR fragments gave rise to 10% low efficiencies compared with those using plasmids.…”