抑制蜜瓜ACC氧化酶的反义RNA-核酶多聚体的研究

Aimed to reduce the biosynthesis of ethylene in melon with antisense technology at RNA level and elongation reserve periods of melon，the genes coding hammerhead ribozyme, that cutting ACC oxidase mRNA of melon ripening periods, and antisense RNA, which inhibit transcription, were designed and chemical synthesized, then the expression vectors in bacterial cell and plant tissues were constructed. In vitro and in vivo experiments of E. coli showed that melon ACC oxidase mRNA was degraded by the antisense RNA- hammerhead ribozyme polymers. The melon Hetao were transformed by expression vectors of antisense RNA- hammerhead ribozyme polymers and three lot of transgenic melon plants which confirmed integrated by Southern blotting were obtained, further examination of the relationship between ACC oxidase activities, ethylene release level and elongation of melon reserve periods.

以河套蜜瓜成熟期ＡＣＣ酶ｍＲＮＡ为靶子序列，拟合成反义ＲＮＡ－核酶多聚体基因，在细菌体内和体外实验证明专一性抑制该靶子序列的基础上，构建诱导型和组成型植物表达载体，转化蜜瓜。在转基因蜜瓜中研究反义ＲＮＡ－核酶多聚体基因按时空调控表达的机制和条件，企图抑制和减缓乙烯的合成和释放，为培育保鲜期较长的蜜瓜品系打下基础。