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KMID : 0903519930360060562
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1993 Volume.36 No. 6 p.562 ~ p.566
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Electrofusion and preparation of transgenic plant by direct insert of marker gene
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유장걸/U, Zang Kual
소인섭/홍경애/류기중/김양록/So, In Sup/Hong, Kyung Ae/Riu, Ki Jung/Kim, Yang Lok
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Abstract
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The conditions required for plant transformation through the electroporation system and for the electrofusion of the prtoplasts were investigated for geranium (Pelargonium zonale hybrids). The optimum condition for electroporation was 1.77 ㎸/㎝ for 40 μsec under which 70% of the protoplasts were viable and 58% of the viable protoplasts were stained with methylene blue. The pBin19 DNA plasmid used as a carrier vector was isolated from E.coli DH5α strain, purified, identified by the electrophoresis on agarose gel and electroporated into the protoplasts. The KM8 liquid medium gave better cell division than any other media. One ㎒ of AC frequency with 40 V/㎝ of amplitude for 15 sec followed by 0.5 ㎸/㎝ of DC amplitude for 60 μsec was most efficient for the electrofusion of protoplasts.
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