An episomal vector system for plastid transformation in higher plants = 고등식물의 엽록체 형질전환을 위한 episomal vector 시스템

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An episomal vector system for plastid transformation in higher plants = 고등식물의 엽록체 형질전환을 위한 episomal vector 시스템
Sung Ran Min; S J Davarpanah; S H Jung; Y I Park; Jang Ryol Liu; Won Joong Jeong
Bibliographic Citation
Plant Biotechnology Reports, vol. 9, no. 6, pp. 443-449
Publication Year
We developed a new plastid transformation vector system using the putative replication origin of a minicircular chromosome from the marine dinoflagellate Heterocapsatriquetra. Transplastomic tobacco plants generated with this vector properly expressed the green fluorescent protein (GFP) gene without incorporating it into the plastid genome. To construct the episomal vector, a 610-bp DNA fragment containing the putative replication origin was fused to a dicistronic expression cassette encoding the aminoglycoside 3′-adenyltransferase (aadA) and gfp genes under control of the plastid rrn promoter. The vector was delivered to plastids of tobacco leaf explants by biolistic bombardment. After 8 weeks of bombardment, episomal transformant shoots were generated from leaf explants cultured on selection media containing 500 mg/L spectinomycin. Fluorescence microscopy and northern blot analysis demonstrated GFP expression in episomal transformant plants. PCR, Southern blot analysis, recovery of episomes, and sequencing analysis showed the vector to be maintained as self-replicating extrachromosomal circular DNA molecules for at least 6 months. Using a single construct for all plants, our episomal vector system may offer an advantage over the conventional plastid vector systems, which require species-specific constructs.
EpisomeHeterocapsatriquetraPlastid transformationTobacco
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Division of Research on National Challenges > Plant Systems Engineering Research > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > Cell Factory Research Center > 1. Journal Articles
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