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By 陳立德, 曾夢蛟, Li-Te Chen and Menq-Jiau Tseng


In this study, bt and hsc70 genes as well as sod 62 and cat 78 genes were constructed in the same Brassica-specific-plastid vector which were named pASCCBtHSC and pASCCSODCAT, respectively. The constructed vectors were transferred into cabbage (KY-cross) chloroplast via particle bombardment mediated transformation. The results of PCR, Southern and Northern blot hybridization indicated that seven plants of co-transformed sod 62+ cat 78 plants contained both sod 62 and cat 78 genes, and expressed sod 62 and cat78 mRNA. One of the two co-transformed bt + hsc 70 plants contained both bt and hsc 70 genes, and exhibited the high degree of resistance to the Plutella xylostella.本研究將熱休克蛋白(heat shock proteins cognate, HSC)基因與蘇力菌殺蟲晶體蛋白 (Bt) 基因構築再同一個蕓苔屬蔬菜葉綠體轉殖載體,稱之為pASCCBtHSC;將超氧化歧化酵素(superoxide dismutase, SOD)基因與過氧化氫酵素(catalase, CAT)基因構築到同一個蕓苔屬蔬菜葉綠體轉殖載體,稱之為 pASCCSODCAT。利用基因槍法將兩種載體分別轉殖到'初秋'甘藍葉綠體中。以PCR、南方墨點與北方墨點分析轉殖再生植株,在 pASCCSODCAT (cat+ sod)方面,獲得的七株轉殖甘藍,均同時具有 cat 及 sod 基因並轉錄出其 mRNA;在 pASCCBtHSC (bt+hsc)方面,獲得的兩株轉殖甘藍中,即有一株轉殖甘藍同時具有 bt 及 hsc70 基因,轉錄出 bt 及 hsc70 mRNA,並大量表現HSC70蛋白及具有顯著的抗蟲效果

Topics: 葉綠體基因轉殖, 甘藍, 基因槍, biolistic bombardment, chloroplast trandgenic, cabbage
Year: 2014
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