Yin Chang-Cheng, Wang Shang-Zi, Lin Zheng-Hong, Wang Xiang-Bin, Liu Jing, Huang Hua-Liang
Institute of Genetics and Developmental Biology, the Chinese Academy of Sciences, Beijing 100101, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Oct;35(10):921-4.
During the construction of a random peptide repertoire using degenerate models, unexpected amino acids or stop codons are almost unavoidable. To conquer this shortcoming, a new split-mix-split method of oligonucleotide synthesis was developed. A 13-amino acids random peptide library had been constructed by using this method. The sequencing results of 16 clones indicated that neither stop codon nor codon for cysteine appeared as designed. The occurrence rations of 19 amino acids were also calculated and no obvious amino acid bias had been observed. By using this method, the type and quantity of amino acid at certain position of a peptide could be controlled well, so this split-mix-split method, combined with degenerate could meet the needs of a high diversity random peptide library.
在使用简并模型构建随机肽文库的过程中,意外的氨基酸或终止密码子几乎是不可避免的。为克服这一缺点,开发了一种新的寡核苷酸合成的拆分-混合-拆分方法。利用该方法构建了一个含有13个氨基酸的随机肽文库。对16个克隆的测序结果表明,既没有出现终止密码子,也没有按设计出现半胱氨酸密码子。还计算了19种氨基酸的出现比例,未观察到明显的氨基酸偏好。通过该方法,可以很好地控制肽特定位置的氨基酸类型和数量,因此这种拆分-混合-拆分方法与简并相结合能够满足高多样性随机肽文库的需求。