1. DNA複製的步驟：
A. 辨認起始點(Ori)，解螺旋酶(Helicases)接上
B. 解螺旋酶(Helicases)：使用將三磷酸腺苷(ATP)水解(hydrolysis)，轉成能量，解開雙股去氧核醣核酸(DNA)
C. 複製叉（replication fork）形成，RNA引子( RNA primer)接上
D. DNA polymerase(DNA聚合酶)會開始進行DNA的合成與延長(elongation)
E. 新合成的DNA片段連接形成複製泡(replication bubbles)
2. 新股複製的方向是由5’端到3’，因此複製時會產生兩種股線：
A. Leading strand(領先股)：根據3’端到5’端的模板股複製，複製出一段由5’端到3’端連續的DNA
B. Lagging strand(落後股)：其模板股的方向為5’端到3’端，所以只好一段段來做(注意：DNA合成方向仍是由5’端做到3’端)，形成岡崎片段，最後會再接起來
3. DNA複製所需之蛋白質：
蛋白質
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功能
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負責DNA的延長，接上去氧核醣核苷酸
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Helicases(解旋酶)
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將雙股DNA解開
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當解旋酶解開雙股DNA時，會產生螺旋壓力，拓樸酶可藉由解開一個環減少螺旋壓力
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DNA primase(DNA引子酶)
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合成RNA引子
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SSB(Single-stranded DNA-binding protein；單鏈DNA-結合蛋白)
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防止兩條互補單鏈再次結合為雙股，維持單鏈狀態，以利DNA合成
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把兩條DNA片段黏合成一條
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一、原核生物(prokaryotes)
1. DNA複製的步驟
A、 DnaA辨識起始點後和DNA做結合
B、 DnaB (helicase)將雙股DNA打開變成單股，SSB蛋白會接到單股上
C、 DNA primase製造RNA primer
D、 RNA primer附著到DNA上後，就開始進行Leading strand和Lagging strand的DNA合成，兩股的合成皆須DNA polymerase III
E、 複製叉後方有DNA topoisomerase幫忙負擔解開螺旋的壓力
1. DNA複製的步驟
A、 DnaA辨識起始點後和DNA做結合
B、 DnaB (helicase)將雙股DNA打開變成單股，SSB蛋白會接到單股上
C、 DNA primase製造RNA primer
D、 RNA primer附著到DNA上後，就開始進行Leading strand和Lagging strand的DNA合成，兩股的合成皆須DNA polymerase III
E、 複製叉後方有DNA topoisomerase幫忙負擔解開螺旋的壓力
2. 落後股的DNA複製
A、 岡崎片段中會有很多的RNA primer，所以落後股的結合需要去除這些RNA primer
B、 DNA polymerase I會利用其5'～3' exonuclease(外切酶)次單元一邊將RNA primer移除，一邊接上去氧核苷酸來補齊空缺的序列
C、 利用DNA ligase把岡崎片段連接起來
3. 原核細胞中不同DNA polymerases的比較
A、 在E.coli裡面，除了DNA polymerase I和DNA polymerase III，還有DNA polymerase II
B、 Polymerase I、II、III，主要分別從polA、polB、polC這三個不同的基因製造出來的
C、 DNA polymerase I、II、III都有從3'～5' Exonuclease的作用，但5'～3' Exonuclease的作用只有DNA polymerase I才有，也因此只有DNA polymerase I才能移除primer
＊DNA polymerase I只能一個一個移除鹼基；DNA polymerase III可以從5'～3' 整段移除
＊DNA polymerase I只能一個一個移除鹼基；DNA polymerase III可以從5'～3' 整段移除
二、真核生物(eukaryotes)
1. 真核生物的DNA複製和原核生物基本上是一樣的，但哺乳類動物卻多了Sliding clamps蛋白的作用
1. 真核生物的DNA複製和原核生物基本上是一樣的，但哺乳類動物卻多了Sliding clamps蛋白的作用
參見：滑動夾子蛋白(Sliding clamps proteins)
2. 複製的步驟：(舉落後股為例)
A、 Step1：RPA蛋白作用為SSB蛋白，結合單股DNA
B、 Step2：剛開始時，利用DNA Polymerase α和RNA primase，先製造出一段RNA primer，然後DNA Polymerase α會製造出一段長約30個核甘酸的DNA序列，之後DNA Polymerase α和primase兩者便會分離。
C、 Step3：RFC(replication factor C，clamp-loading factor)會結合到DNA上，會組合出PCNA(proliferating cell nuclear antigen,作為sliding clamp)，夾住DNA。之後RFC便會離開
D、 Step4：DNA Polymerase δ會和PCNA結合，一起進行DNA的複製
E、 Step5：在即將碰到後面那段岡崎片段時，RNaseH會附著到RNA引子上負責將其上的RNA核苷酸移除，再用FEN1移除最後一個核苷酸。
c.f.比較：原核生物則是用DNA PolymeraseⅠ將RNA primer移除掉
F、 Step6：PCNA和DNA Polymeraseδ繼續合成DNA補足剛剛RNA引子所在位置的序列，最後用DNA ligase將片段連接起來
2. 複製的步驟：(舉落後股為例)
A、 Step1：RPA蛋白作用為SSB蛋白，結合單股DNA
B、 Step2：剛開始時，利用DNA Polymerase α和RNA primase，先製造出一段RNA primer，然後DNA Polymerase α會製造出一段長約30個核甘酸的DNA序列，之後DNA Polymerase α和primase兩者便會分離。
C、 Step3：RFC(replication factor C，clamp-loading factor)會結合到DNA上，會組合出PCNA(proliferating cell nuclear antigen,作為sliding clamp)，夾住DNA。之後RFC便會離開
D、 Step4：DNA Polymerase δ會和PCNA結合，一起進行DNA的複製
E、 Step5：在即將碰到後面那段岡崎片段時，RNaseH會附著到RNA引子上負責將其上的RNA核苷酸移除，再用FEN1移除最後一個核苷酸。
c.f.比較：原核生物則是用DNA PolymeraseⅠ將RNA primer移除掉
F、 Step6：PCNA和DNA Polymeraseδ繼續合成DNA補足剛剛RNA引子所在位置的序列，最後用DNA ligase將片段連接起來
資料來源：維基百科
3. 真核生物(eukaryotes)具有多個起始點：
A、 真核生物同一條DNA上具有多個複製起始點串聯在一起(tandem replicons；串聯式複製體)；而原核生物卻只有一個起始點
B、 在真核生物中，DNA複製是雙向的(bidirectional)，也就是說在每一個起始點DNA可以朝兩個方向進行複製
C、 複製完後再把所有片段連接在一起，由於多個複製起始點同時開始複製基因，因此可以快速複製完真核生物的DNA
▼遺傳物質(genetic material)
顯示/隱藏(show/hide)
二、"真"核生物(eukaryotes) <<< 有錯誤!
回覆刪除感謝您的更正!!!
刪除歡迎再次來訪
雙向的(biderectional)，Bidirectional
回覆刪除感謝您的更正!!!
刪除歡迎再次來訪
應該是 Leading strand
回覆刪除感謝您的更正!!!
刪除歡迎再次來訪
不好意思想請問
回覆刪除在真核生物那邊1.提到
真核生物的DNA複製和原核生物基本上是一樣的，但哺乳類動物卻多了Sliding clamps蛋白的作用
但是點進下方連結後裡面又提到
不只真核生物具有滑動夾子蛋白(Sliding clamps proteins)，其他的原核生物、古細菌、病毒都有
不知道是哪邊有筆誤或是其中有細節是我不了解的部分
求解感謝~
原核生物裡應該是beta-clamp(polymerase III holoenzyme的beta subunit)
刪除原核跟真核都有sliding clamp歐！！！內文敘述是不是有誤…？
回覆刪除DNA複製完還會在加工嗎?
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C、 DNA polymerase I、II、III都有從3'～5' Exonuclease的作用，但5'～3' Exonuclease的作用只有DNA polymerase I才有，也因此只有DNA polymerase I才能移除primer
回覆刪除＊DNA polymerase I只能一個一個移除鹼基；DNA polymerase III可以從5'～3' 整段移除
不是說''5'～3' Exonuclease的作用只有DNA polymerase I才有''，怎麼最後一句又寫DNA polymerase III可以從5'～3' 整段移除，求解釋~~
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請問:何謂poly(A) RNA?
回覆刪除想請問真核細胞，DNA複製時RNA引子是由誰合成的？
回覆刪除我閱讀的書中提到DNA polymerase α中就已經有primase 的次單元了，還需要RNA polymerase嗎