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Electroporation 에 의한 담배 세포로의 유전자 도입
이광웅,이종섭,서정원 한국유전학회 1989 Genes & Genomics Vol.11 No.2
In order to improve the method of plant transformation, transfer of DNA into intact cells of tobacco was carried out by electroporation. Chimaeric genes between tomato proteinase inhibitor I and chloramphenicol acetyltransferase genes were constructed in a vector derived from Ti-plasmid of Agrobacterium tumefaciens. Suspension cells of tobacco were electric-shocked in a chamber with DNAs of one of the constructs, pJSL105 and were plated on culture medium containing kanamycin. When shocked at 4㎸/㎝, colonies were formed from the cells on the medium containing 200㎍/ml kanamycin. The kanamycin-resistant colonies were further cultured to form calluses, which were regenerated into shoots. Roots were initiated from the shoots derived from kanamycin-resistant calluses while not from control. Southern hybridization of genomic DNAs isolated from leaves of a transgenic plant revealed the presence of the plasmid DNA. This indicated that intact cells can be used as plant material for the gene transfer by electroporation.