葉綠體在植物的生長發育過程中扮演重要的角色。然而葉綠體基因組容易受到紫外線以及光合作用電子傳遞過程所產生的活性氧化物而傷害，導致植物的生長發育受抑制。因此，葉綠體必須有能夠修復受損基因組之方法，以維持基因組的穩定。其中同源重組是一種能完整修復受損基因組的方式。實驗室的學姐得到轉殖Transcription activator-like effector nuclease (TALEN)基因之阿拉伯芥，其作用標的為葉綠體的rpoB基因。本研究將轉殖株進行篩選直到獲得同型合子之植株。利用表現TALEN的轉殖植株與會造成胞器DNA雙股斷裂之環丙沙星(ciprofloxacin)處理植株並且比較涉及同源重組之基因表現量的變化。結果顯示表現TALEN蛋白的轉殖株會對涉及葉綠體DNA同源重組之基因的表現造成影響，但是與環丙沙星處理的植株比較，發現基因表現量的變化模式不一致，推測環丙沙星對細胞造成更廣泛且嚴重的影響，而造成修復機制策略的差異。而在表現TALEN蛋白的轉殖株，其促進同源重組過程相關的基因如ODB2、WHY3、OSB3、RECA1和MSH1等基因之表現量下降，顯示可能轉殖株以小片段同源重組的方式為主要途徑進行修復，但還需要後續實驗之驗證。

Chloroplast is the place for photosynthesis to occur. The chloroplast DNA (cpDNA) might be damaged by UV and reactive oxygen species producing through electron transport chain. The DNA double-stranded break (DSB) is the most severe damage for cpDNA. To maintain the genome stability, the chloroplast must confer some strategies to repair the damaged genome. Among them, homologous recombination (HR) is a process that can completely repair damaged DNA. Previously, transgenic Arabidopsis with the overexpression of transcription activator-like effector nuclease (TALEN) driven by two different promoters, rbcS and β-estrodiol-inducible promoters, was generated in the Lab. The recombinant TALEN protein is expected to specifically bind to rpoB gene and cause the DSB of cpDNA. In this study, transgenic plants were selected until the homozygous stage and was used to study the expression of genes involved in the DSB repairing through HR by qRT-PCR assay, and compared with that of plants treated with ciprofloxacin, which it conferred no specificity, but was commonly used to cause DNA DSB in mitochondrial and chloroplast. According to the preliminary result, the relative expression level of most genes involved in the HR process was altered in the TALEN transgenic plants. But the relative expression level of most genes in TALEN transgenic plants was not consistent with that of plants treated with ciprofloxacin. It was speculated that ciprofloxacin might cause more extensive and severe effects on the DNA damage of plant cells. However, in TALEN transgenic plant, the decrease in the expression level of genes such as ODB2、WHY3、OSB3、RECA1 and MSH1 might suggest that microhomology-mediated end-joining (MMEJ) is the major repairing process for the DSB of cpDNA in transgenic Arabidopsis. However, further research is needed to verify the hypothesis.

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