DNA偶合物构建思路: Difference between revisions
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Revision as of 15:47, 8 October 2025
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protein
gene engineering and click chemistry
- Click Chemistry:通过叠氮基团和炔基(环炔)环加成生成三唑环结构

- 构建生物正交系统:
1.在目标蛋白末端基因(N端)插入琥珀密码子(TAG-终止密码子)
2.在培养系统中引入特定的tRNA(特异性识别TAG位点)与其对应的氨酰-tRNA合成酶(能将p-Azidophenylalanine(含有一个叠氮基团)这个非天然氨基酸连接到tRNA上)
3.BCN-DNA(BNN含环炔)

zinc finger protein(zif268)
- zif268:含三个“锌指”模块,可插入连续的三个大沟(3×3)中,识别特定的基因序列(例如:5'-GCG TGG GCG-3')

the basic leucine-zipper protein(GCN4)
- GCN4:通过形成二聚体,像夹子一样夹住双螺旋两侧的大沟中(通常和目标蛋白通过基因工程用连接子连接)

SNAP-tag
- SNAP-tag:一种蛋白质标签,可与底物 benzylguanine (BG) 发生共价结合,BG通常可用被修饰在T上面