title: TRANSIENT TRANSFORMATION OF A CHLORARACHNIOPHYTE ALGA, LOTHARELLA AMOEBIFORMIS (CHLORARACHNIOPHYCEAE), WITH uidA AND egfp REPORTER GENES(1).
abstract: A transient genetic transformation system was established for a chlorarachniophyte alga, Lotharella amoebiformis K. Ishida et Y. Hara. We first isolated sequences that contain a putative promoter for a RUBISCO SSU (rbcS) gene and a terminator for another copy of rbcS gene from L. amoebiformis. With those promoter and terminator sequences, we developed two expression vectors, pLaRGus and pLaRGfp, which code uidA and egfp genes, respectively. The cells were then transformed with each vector using a microparticle bombardment system. When the cells were transformed with the pLaRGus, β-glucuronidase (GUS) staining dyed several cells blue. Green fluorescent protein (GFP) fluorescence was observed in the cells transformed with pLaRGfp. The highest transient transformation efficiency, 35 per 2 × 10(7) cells, was detected from the GUS staining. This study demonstrates that two reporter genes are expressed in L. amoebiformis cells when rbcS promoter and terminator are used. The conditions of transformation were also optimized. This is the first report of successful genetic transformation in chlorarachniophyte algae.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, JapanDivision of Life Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa 920-1192, JapanGraduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Hirakawa
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, JapanDivision of Life Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa 920-1192, JapanGraduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, Japan.
authors.forename: Rumiko
authors.initials: R
authors.lastname: Kofuji
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, JapanDivision of Life Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma, Kanazawa 920-1192, JapanGraduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1, Tennodai, Tsukuba 305-8572, Japan.
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Ishida
keywords: GFP; GUS; chlorarachniophyte; microparticle bombardment; transient transformation