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構築基因至同一個葉綠基因體轉殖載體以基因槍法共同轉殖基因到甘藍葉綠體

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

Abstract

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
OAI identifier: oai:ir.lib.nchu.edu.tw:11455/81787
Journal:

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