基因槍轟擊法是葉綠體基因改造中最主要使用的技術，可將表現載體送入葉綠體中，再藉由篩選與在生來獲得葉綠體轉殖植物。然而，以基因槍轟擊法來獲得轉殖植物花費高，且篩選與在生的過程相當耗時。因此本研究應用 Transcription activator-like effectors nucleases (TALENs) 之基因編輯技術特異性修飾阿拉伯芥葉綠體編碼的 rpoB 基因。構築兩組以不同啟動子表現重組 TALEN 基因的表現載體，分別是由 rbcS 啟動子來調控基因表現，另一組為由雌激素誘導表現的啟動子調控基因表現，利用農桿菌媒介花序法轉殖阿拉伯芥，已獲得7株由 rbcS 啟動子來調控基因表現的T0 阿拉伯芥轉殖株 (RbcS-M 品系)，並且以 PCR 分析可以同時偵測到 TALEN-L 與 TALEN-R 外源基因的存在。已獲得3株由雌激素誘導表現的啟動子調控基因表現的T0 阿拉伯芥轉殖株 (pER8-M 品系)，並且以 PCR 分析可以同時偵測到 TALEN-L 與 TALEN-R 外源基因的存在。然而，以西方墨點轉印法分析，目前無法檢測到TALEN 蛋白的表達。但在外表性狀的觀察中，一些轉基因的阿拉伯芥轉殖株在早期生長階段顯示與未轉基因擬南芥的表型差異。

The major method of the plastid DNA modify was the biolistic bombardment delivering expression vectors into plastid. The transplastomic plants were obtained by selection and regeneration process. However, biolistic bombardment to obtain the transplastomic plants was costly, and selection and regeneration process was time-consuming. In this study, apply transcription activator like effector nuclease (TALENs) editing technology to site-specifically modify the plastid rpoB gene. Two pairs of nuclear TALENs expression vectors, one driven by rbcS promoter, the other driven by estrogen-inducible promoter. Agrobacteria-mediated floral dip method was applied to transform Arabidopsis. Seven rbcS promoter-regulated transgenic Arabidopsis lines (RbcS-M) which PCR test positive were obtained. Three chemical-inducible promoter regulated transgenic Arabidopsis lines which PCR test positive were obtained. However, the expression of TALEN proteins was not detectable by Western blot analysis so far. Some transgenic Arabidopsis lines showed phenotypic difference with untransgenic Arabidopsis in early growth stage.

王紫萱、易自力，卡那霉素在植物轉基因中的應用及其抗性基因的生物安全性評價，中國生物工程雜誌，9-13，2003。

涂晉敏，應用創傷弧菌藍螢光蛋白於植物科學研究，國立成功大學熱帶植物科學研究所碩士論文，2014。

蘇源霖，轉殖似轉錄激活因子蛋白和酸酶 (TALEN) 來改變菸草葉綠體 DNA 序列之研究，國立成功大學生物科技研究所碩士論文，2016。

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