[논문]Agrobacterium-Mediated Transformation System for Large-Scale Producion of Transgenic Chinese Cabbage (Brassica rapa L. ssp. pekinensis) Plants for Insertional Mutagenesis

Lee, Mi-Kyung; Kim, Hyoung-Seok; Kim, Jung-Sun; Kim, Sung-Hoon; Park, Young-Doo

doi:10.1007/bf03030544

Agrobacterium-Mediated Transformation System for Large-Scale Producion of Transgenic Chinese Cabbage (Brassica rapa L. ssp. pekinensis) Plants for Insertional Mutagenesis

Journal of plant biology = 식물학회지, v.47 no.4, 2004년, pp.300 - 306

Lee, Mi-Kyung (Graduate School of Biotechnology and Department of Horticultural Biotechnology, KyungHee University) , Kim, Hyoung-Seok (Graduate School of Biotechnology and Department of Horticultural Biotechnology, KyungHee University) , Kim, Jung-Sun (Brassica Genomics Team, National Institute of Agricultural Biotechnology) , Kim, Sung-Hoon (Graduate School of East-West Medical Science, KyungHee University) , Park, Young-Doo (Graduate School of Biotechnology and Department of Horticultural Biotechnology, KyungHee University)

Abstract ▼ AI-Helper

In order to better utilize insertional mutagenesis and functional genomics in Chinese cabbage, we have developed an improved transformation system that more efficiently produces a large number of transgenic plants. Hypocotyl explants were inoculated with Agrobacterium tumefaciens LBA4404. This strain harbors tagging vector pRCV2, which contains a hygromycin-resistance gene, an ampicillin resistance gene, and a bacterial replication origin within the T-DNA. Transformation efficiency was highest when the explants were first co-cultivated for 3 d in a medium supplemented with 5 mg L-1 acetosyringone, then transferred to a 0.8% agar selection medium containing 10 mg L-1 hygromycin. In addition, maintaining a low pH in the co-cultivation medium was critical to enhancing transformation frequency. A total of 3369 transgenic plants were obtained, with efficiencies ranging from 2.89% to 5.00%. Southern blot analysis and T1 progeny tests from 120 transgenic plants confirmed that the transgenes were stably inherited to the next generation. We also conducted plasmid rescue and inverse PCR with some transformants, based on their phenotype, to demonstrate the applicability of T-DNA tagging in Chinese cabbage. The tagged sequences were then analyzed.

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참고문헌 (20)

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저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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