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식물배양세포의 저장관계 유전자탐색과 형질전환에의 활용 1. 벼 배양세포의 건조보존기구와 관련유전자 선발
김길웅,대야청춘,신동현,Virigool, S.,Shinzaki, K. Y.,홍석영 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
A system for long-term dry preservation of rice (Oryza sativa L.) callus and a protection mechanism regulating the survival of dried callus were investigated. The highest survival of dried callus and the highest regeneration of plantlets were observed in calli which had been pretreated with 10^(-5) M abscisic acid (ABA) in the presence of 90 g/L of sucrose. A corresponding accumulation of the RNA of the rab 16A gene (a rice gene induced by ABA and water stress) was detected in dried callus, mature seeds, and callus pretreated with 10^(-5) M ABA. Analysis of protein by two dimensional polyacrylamide gel electrophoresis(2D-PAGE) demonstrated different protein patterns in dried callus pretreated with 10-s M ABA and 90 g/L of sucrose compared to callus dried without the pretreatment. Three different cDNA clones, pDHS1, pDHS2, and pDHS3 contained cDNA inserts in size of 3.8, 3.2, and 3.2 kilobase pairs, respectively, were isolated by using Nhe I fragment from rab 16A as a probe. All the three clones exhibited unique restriction maps.