Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/81718
標題: Co-transformation of Bt-toxin, Heat Shock Protein, Superoxide Dismutase, and Catalase Genes into Cabbage(Brassica oleracea L. var. capitata L.) via Agrobacterium Mediated Transformation
共同轉殖蘇力菌殺蟲結晶蛋白、熱休克蛋白、超氧化歧化酵素及過氧化氫酵素基因至甘藍之研究
作者: 葉元純
Yuan-Chun Yeh
曾夢蛟
Menq-Jiau Tseng
關鍵字: 過氧化氫酵素
Co-transformation
熱休克蛋白
超氧化歧化酵素
蘇力菌殺蟲晶體蛋白
農桿菌共同轉移法
Catalase
Superoxide Dismutase
Heat Shock Protein
Cabbage
Bt-toxin
摘要: Attempts had been made to co-transfer the Bt-toxin gene (bt), superoxide dismutase gene (sod62) of Chinese cabbage, catalase gene (cat78) of Chinese cabbage, and heat shock protein gene (hsc70) of tomato into the cabbage (Brassica oleracea L. var. K-Y cross). The objectivesof this study are to establish the technology of gene co-transferand targeting the foreign proteins into the chloroplast by chloroplast trainsit peptide, and to study the possibility for improvement of cabbage with insect, disease, and stresses resistance by genes co-trans-formation. The results of PCR, Southern nad Northern hybridization indicated that the bt, hsc70, sod62 and cat78 genes had been inserted into the genome of cabbage, and expressed. The result of isozyme profiles assay revealed that the transformed SOD62 and CAT78 were present in the transformed plants. The SOD and CAT activities in leaves and chloroplast of transformed plants were higher than those of the controls. Increases in amounts of HSC70 protein were found in the transformed plants s analyzing by western blot hybridization. Preliminary results of bioassay indicated that the increases in tolerance to high temperature (42℃, 4 days), SO2 (800 ppb, 3 days), and insect resistance (Plutella xylostella) were found in the transgenic plants. This study demonstrated that two, three and four foreign genes could be co-transgenic plants. This study demonstrated that two, three and four foreign genes could be co-transferred into a cabbage plant simultaneously via Agrobacterium mediated transformation.
本研究是利用農桿菌共同轉移法,研究同時轉移帶有 rbcS 為啟動子且攜帶有大豆葉綠體訊息 transit peptide 之蘇力菌殺蟲晶體蛋白 (bt)、結球白菜之超氧化歧化酵素 (sod62)、結球白菜之過氧化氫酵素 (cat78) 及番茄之熱休克蛋白 (hsc70) 等四種基因至同一棵之'初秋'甘藍,並探討轉運蛋白質到葉綠體之可行性。 以PCR、南方墨點及北方墨點雜交分析再生之bt、hsc70、sod62及cat78基因共同轉移的甘藍植株,試驗結果顯示轉移之基因已插入到轉殖植株染色體上,並表現出bt、hsc70、sod62及cat78基因之mRNA。電泳膠片酵素活性染色分析之結果顯示,轉殖植林之SOD及CAT條帶,均較未轉殖植株之條帶濃,其SOD及CAT酵素活性之對照組顯著增加。西方墨點分析結果顯示,轉殖植株之HSC70顯著較未轉殖植株之對照組多,且能轉連到葉綠體內。 具有轉殖bt基因之甘藍再生植株葉片經餵食小菜蛾幼蟲,證實具有抗蟲之效果。以800ppb之SO2燻蒸轉殖植株三天,具有sod及cat基因之甘藍再生植株,對於SO2 之危害較未轉殖或只轉殖單一sod或cat基因之轉殖株具有抗性。本研究證實用農桿菌混合感染培殖體可達到多基因同時轉移到同一棵甘藍,且能正常表現有功能之蛋白質,使轉殖植株具有抗蟲、耐高溫及耐逆境之特性。
URI: http://hdl.handle.net/11455/81718
ISSN: 0255-5921
Appears in Collections:第28卷 第03期
園藝學系

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