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標題: 阿拉伯芥中 FYF 可能的複合體研究與AGL6 的功能性分析
Characterization of FYF protein complexes and functional analysis of AGL6 in Arabidopsis
作者: 曹晉瑋
Chin-Wei Tsao
關鍵字: AGL6;AGL6
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FYF 為一個MADS-BOX 蛋白可以藉由促進FUF1 抑制EDF1-4、BOP1/2、
IDA 基因延緩花朵的老化和脫落。為了知道哪些蛋白參與了FYF 所形成的蛋白
四聚體以調控花朵的老化和凋落,利用酵母菌雙雜交系統的篩選找出和FYF 具
有交互作用的候選蛋白。其中發現AGL6 是一可與FYF 進行交互作用的候選蛋
白。為進一步探討AGL6 與花朵的老化和凋落之關係,將35S::AGL6 及
35::AGL6+SRDX 轉殖入阿拉伯芥,並分析轉殖植株之性狀。結果發現在兩種轉
基因阿拉伯芥中都有早開花的情況,此外35S::AGL6+SRDX 可以藉由抑制
EDF1/2、IDA、BOP1 產生延緩花朵老化凋落的性狀。為了進一步證實酵母菌雙
AGL6 可以幫助FYF 進入細胞核,並且兩者間FRET 效率有55%。FRET 的效率
和蛋白之間的距離是成反比的,FRET 效率越高表示兩蛋白之間距離越近。在FYF
與候選基因AGL15 的交互作用中發現,FYF 可與AGL15 和AGL6 穩定形成四
聚體,AGL71 為另外一個FYF 的候選交互作用蛋白,AGL71 也可以與AGL15
和兩份AGL6 形成穩定四聚體。上述之結果顯示AGL71 能夠取代FYF 與AGL6
和AGL15 交互作用形成四聚體。總結來說,當AGL6 扮演著一個抑制子的角色
時和FYF 一樣可以延緩花朵老化脫落。未來將更加了解MADS-Box 蛋白間的交
互作用關係,藉以建立MADS-Box 蛋白調控花朵老化脫落的模型。

FOREVER YOUNG FLOWER (FYF), a MADS-Box gene can delay flower
senescence and abscission by up-regulating FYF UP-REGULATING FACTOR 1 (FUF1)
proteins formed tetramer protein complex with FYF in regulating flower
senescence/abscission, yeast-two-hybrid screen was performed. AGAMOUS-LIKE 6
(AGL6), one of the potential FYF interacting proteins was identified. To further analyze
the function of AGL6 in regulating flower senescence and abscission, 35S::AGL6,
35::AGL6+SRDX transgenic Arabidopsis were generated. Early flowering was
observed in these two different types of transgenic Arabidopsis. In addition,
35S::AGL6+SRDX could delay flower senescence/abscission by repressing EDF1/2,
IDA, BOP1 in transgenic Arabidopsis. The further confirm the interaction between FYF
and AGL6, fluorescence resonance energy transfer (FRET) was used. The result
indicated that AGL6 is able to help FYF to transport into the nucleus and the FRET
efficiency between FYF and AGL6 is about 55%. It is well known that FRET efficiency
and proteins distance are inversely proportional. The higher FRET efficiency indicates
the shorter distance between two proteins. AGL15, the other candidate interaction
protein which could form a protein tetramer with FYF and AGL6. AGL71 also could
form a tetramer with AGL15 and AGL6. The result indicated that AGL71 could replace
FYF to form a tetramer with AGL15 and AGL6. In the future, how MADS-box protein
to form tetramer protein complex in regulating flower senescence and abscission will
be further analyzed.
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