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TRANSIENT TRANSFORMATION OF A CHLORARACHNIOPHYTE ALGA, LOTHARELLA AMOEBIFORMIS (CHLORARACHNIOPHYCEAE), WITH uid A AND egfp REPORTER GENES 1
Author(s) -
Hirakawa Yoshihisa,
Kofuji Rumiko,
Ishida Kenichiro
Publication year - 2008
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.1529-8817.2008.00513.x
Subject(s) - terminator (solar) , biology , green fluorescent protein , transformation (genetics) , reporter gene , gene , microbiology and biotechnology , genetics , gene expression , ionosphere , physics , astronomy
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 uid A 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.