Alanine-stretch scanning mutagenesis: A simple and efficient method to probe protein structure and function
Alanine-stretch scanning mutagenesis: A simple and efficient method to probe protein structure and function
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DOI：
10.1093/nar/25.2.447
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发表时间：
1997-01-15
影响因子：
14.9
通讯作者：
Masson, JM
中科院分区：
文献类型：
--
作者：
Lefevre, F;Remy, MH;Masson, JM
我们已经开发出一种简单的方法，用丙氨酸代替一段残基。在通过IPCR引入PsfI位点，从而产生两个丙氨酸密码子之后，通过使用“丙氨酸-延伸片段”在该位点引入另外的密码子。这些盒包含两侧侧翼为丙氨酸密码子的抗生素抗性基因。然后通过PvuII切除抗性基因产生密码子的正确插入。该方法可靠性高、灵活性强，具有一定的通用性。
We have developed a foolproof method to substitute a stretch of residues by alanines. After the introduction of a PsfI site by IPCR, thus creating two alanine codons, additional codons are introduced at this site through the use of an 'alanine-stretch cartridge'. These cartridges comprise an antibiotic resistance gene flanked on both sides by alanine codons. Excision of the resistance gene by PvuII then yields the correct insertion of codons. The method is both highly reliable and flexible and should be of general use.