title: Heterologous overexpression of sfCherry fluorescent protein inNannochloropsis salina.
abstract: Oleaginous microalgae of theNannochloropsisgenus are considered excellent candidates for biofuels and value-added products owing to their high biomass productivity and lipid content. Here, we report the first overexpression and detection of a heterologous sfCherry fluorescent protein inNannochloropsis salinain order to develop a transformation toolbox for future genetic improvements. Particle bombardment was employed for transformation, and expression of Shbleunder the control of TUB and UEP promoters, cloned fromN. salina, was used to confer resistance to Zeocin antibiotics, resulting in 5.9 and 4.7 transformants per 108cells, respectively. Stable integration of the markers into the genome was confirmed using a restriction enzyme site-directed amplification (RESDA) PCR. The expression of sfCherry fluorescent protein was confirmed by Western blot analysis and confocal microscopy. These results suggest new possibilities of efficient genetic engineering ofNannochloropsisfor the production of biofuels and other biochemicals.
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Nam Kyu
authors.initials: NK
authors.lastname: Kang
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Gang-Guk
authors.initials: GG
authors.lastname: Choi
authors.affiliation: Advanced Biomass R&D Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Eun Kyung
authors.initials: EK
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Sung-Eun
authors.initials: SE
authors.lastname: Shin
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Seungjib
authors.initials: S
authors.lastname: Jeon
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea; Advanced Biomass R&D Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Min S
authors.initials: MS
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Byeong-Ryool
authors.initials: BR
authors.lastname: Jeong
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea; Advanced Biomass R&D Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Yong Keun
authors.initials: YK
authors.lastname: Chang
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea; Advanced Biomass R&D Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Ji-Won
authors.initials: JW
authors.lastname: Yang
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Bongsoo
authors.initials: B
authors.lastname: Lee
keywords: EPA, eicosapentaenoic acid; Heterologous protein expression; MARs, matrix attachment regions; Particle bombardment; RESDA PCR, restriction enzyme site-directed amplification polymerase chain reaction; Restriction enzyme site-directed amplification (RESDA) PCR; TAIL PCR, thermal asymmetric interlaced PCR; sfCherry fluorescent protein