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배추에서 삽입돌연변이체 유기를 위한 Ac/Ds 전이인자 시스템의 도입
유재경(Jae-Gyeong Yu),이미경(Mi-Kyung Lee),우은택(Eun-Taek Woo),엄석현(Seok Hyun Eom),김정선(Jung-Sun Kim),박영두(Young-Doo Park) 한국원예학회 2010 원예과학기술지 Vol.28 No.2
Insertional mutagenesis has proven to be a useful method for the isolation of genes defined by a mutant phenotype. Now Activator/Dissociation (Ac/Ds) transposon system has become a very popular tool for gene tagging and functional genomics in various plant species. This study was carried out to produce a large number of insertional mutant lines by using Ac/Ds transposon system. To apply Ac/Ds transposon system, hypocotyl explants of Chinese cabbage ‘CT001’ (inbred line, Carrotop Seed Co., Korea) were transformed by Agrobacterium-mediated transformation method using Agrobacterium GV3101 strain harboring the activation tagging T-DNA vector, pJJ91. Thirty three pJJ91 transgenic plants were confirmed by PCR and Southern blot analysis. Then, the finally selected 23 lines were treated with four different levels (2, 5, 10, and 15 ㎛) of dexamethasone (DEX) to activate Ac and to transpose Ds by spraying to stigma at fertilization stage. We established that the pollination after 5 or 10 ㎛ DEX spray was an appropriate treatment condition for Ds transposition in Chinese cabbage transgenic plant with pJJ91 vector. The 340 bp PCR product that was the expected size after transposition of Ds fragment in T1-progeny was shown and sequenced. Southern blot analysis of T1 transgenic plants was conducted to analyze the possibility of Ds transposition under DEX treatments. From PCR and Southern blot analysis, 8 of 23 lines showed Ds transpositions to other sites of genome. These results mean that Ac/Ds transposon system could be applied to Chinese cabbage for production of large scale insertional mutant lines.