Study On The Construction Of A Plant Expression Vector With Aco1 Antisense Gene Regulated By Fruit Specificity Promoter And Its Transformation To Cucumis Melon 'yu Jingxiang'

Cucumis melon is widely cultivated throughout the world, it has importance economic value to grow. But it is easy to rot in the storage and transport processIn recent years, a set of transgene plants was acquired by the using of genetic engineer for the controlling of ripening in melon. However, the great majority methods adopt the 35S promoter to control the Expression Vector. Although gene can be expression stable, but non-specific expression will result in energy waste, the expression of foreign genes affected.In this study, it is focus on the waste of the energy and the security of the antibiotic caused by the using of the constitutive promoter in the ACC oxidase in the transgenic melon. Based on the cloned CM-ACO1 promoter, following researches have been done:1. pCB-aACO1 was digested with restriction endonuclease HindⅢand Xbar I,and pCM-ACO1-promoter was digested with restriction endonuclease HindⅢand Bam H I.Then use Klenow Fragment end up flat.Becase the promoter was modified by the Hind III and BamHI, the acquired target genes fragment, digested by the restriction endonuclease, was matched to the wirelike vector. And finally, a plant expression vector named pCB-aACO1-P with NPTIIgene was constructed.2. The ACO1-promoter was transferred into Agrobacterium tumefacien EHA105.The Agrobacterium tumefacien was transferred into the plants through the Kan resisting screening and PCR. The transference with the Agrobacterium tumefacien to Nicotiana tabacum were further confirmed the feasibility of the vector.3. Choosing melon cultivated in MS+2mg/L 6-BA, the rate of adventitious buds has 86.12%.Then extend cultivated in MS+0.1mg/L 6-BA and rhizogenic cultivated in 1/4MS. The 5d of Cotyledon leaf age is better for callus4. Adopt optimized transformation system of the melon cultivar and co-cultured for 3days; 200 explants were transformed, and 12 transformed plants, with Kana resisting and masculine PCR were acquired. Some of these plants were cultivated in the greenhouse and the situation of growing were observe.