[An efficient approach to the synthesis of a high-quality random peptide repertoire].

Chang-Cheng Yin, Shang-Zi Wang, Zheng-Hong Lin, Xiang-Bin Wang, Jing Liu, Hua-Liang Huang
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Abstract

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.

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一种合成高质量随机肽库的有效方法。
在使用退化模型构建随机肽库时,意想不到的氨基酸或停止密码子几乎是不可避免的。为了克服这一缺点,提出了一种新的分裂-混合-分裂合成寡核苷酸的方法。用该方法构建了一个13个氨基酸的随机肽库。16个克隆的测序结果显示,终止密码子和半胱氨酸密码子均未出现。计算了19种氨基酸的出现比,未发现明显的氨基酸偏倚。利用该方法可以很好地控制肽段特定位置氨基酸的种类和数量,因此该方法结合简并可以满足高多样性随机肽库的需要。
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