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Title 

An episomal vector system for plastid transformation in higher plants

 

고등식물의 엽록체 형질전환을 위한 episomal vector 시스템

Authors 

Sung Ran MinS J DavarpanahS H JungY I ParkJang Ryol LiuWon Joong Jeong

Publisher 

Springer Verlag (Germany)

Issue Date 

2015

Citation 

Plant Biotechnology Reports, vol. 9, no. 6, pp. 443-449

Keywords 

EpisomeHeterocapsatriquetraPlastid transformationTobacco

Abstract 

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.

ISSN 

1863-5466

Link 

http://dx.doi.org/10.1007/s11816-015-0381-4

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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