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Studies on the Cd-binding protein, antifreeze protein and Bt- toxin genes transfer into three species of Brassica spp.
small subunit（rbcS）及 alcohol hydrogenase（ adh ）啟動子的轉殖
菜之再生率在 1 - 3% 之間。檢測轉殖植株之 GUS 基因，有 95% 以上呈
正反應。以南方墨點法分析 PCR 正反應之轉殖植株，均可偵測到轉移基
在轉殖植物體內均有轉錄。無論 MT-Ⅱ、AF 或 Bt 基因以rbcS啟動子質
體的轉殖後再生植株，其 RNA 表現量較轉殖以 CaMV 35S 啟動子質體的
植株高。經 Bt 基因轉移之植株的生物檢定結果顯示，以 rbcS 啟動子為
轉殖質體的質株，殺小菜蛾效果較以 CaMV 35S 啟動子為轉殖質體的植株
This project includes construction of MT binding protein genes from Geaniue pig (MTⅡ), antifreezing protein genes from winter floumder(AF), and insecticidal protein genes seperated from Bacillus thuringiensis (Bt) with vector carrying with rubisco small subumit (rbcS) and alcohol dehydrogenase (adh) promoter sites respectively. Constructed DNAs were transferred into hypocotyls and cotyledons of three Brassica vegetables on agrobacterium-mediated base. We have focused on establishing gene transter and regeneration systems on Broccoli, Cauliflower and Pak-choi, comparisons in gene expression led by different promoters. Also, assessments on the possibilities of vegetables that are resistant to Cadmium, freezing, and Plutella xylostella are made during discussion. Results have shown that the regeneration rate of five transgenic species among three Brassica vegetables ranges around 1-3%, more than 95% of these plants are GUS positive. Transferred genes are detected by southern assay among PCR selected plantlets; transcriptions of the three genes in transgenic plantlets are also confirmed resulting from Northern assay. Plantlets with rbcS promoter perform a higher RNA expression than those with CaMV 35S promoter, no correlations are fond with different genes and their RNA expression. Beside, results also have show that plantlets with rbcS promoter get higher mortalities of Plutella xylostella rather than those with CaMV 35S promoter which are evidenced by Bt bioassay. Regenerations of three Brassica vegetables with three distinct genes have been observed nowadays, among which the Bt transgenic plantlets have complete bioassays,the other AF and MTⅡ transgenic plants have also exhibit transgenic mRNA that need further bioassays.
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