一种构建高质量随机肽库的有效方法  

An Efficient Approach to the Synthesis of a High-quality Random Peptide Repertoire

作　　者：尹长城[1] 王尚资[1] 林正红[1] 王祥斌[1] 刘晶[1] 黄华樑[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《生物化学与生物物理学报》2003年第10期921-924,共4页

基　　金：国家高技术研究发展计划 ( 863计划 )项目资助 (No .2 0 0 1AA2 15 3 81)~~

摘　　要：为构建完全随机化的基因工程肽库 ,克服现有简并方案中终止密码子和序列组成偏歧的不足 ,提出了一种新的简并DNA文库合成方式。通过这种分组式合成方式构建的肽库可以避免终止密码子的出现和氨基酸组成偏歧的发生 ,还可以控制随机化过程中不同氨基酸的参入比例。以一个 13肽库的合成过程为例对分组式合成法进行了实验 ,测序结果和对 19种氨基酸出现频率的统计表明没有终止密码子和半胱氨酸密码子出现 ,各氨基酸的出现频率接近均值 ,表明这种分组 混合 分组 ,辅以简并合成的方法是行之有效的 ,能满足各类高容量基因工程随机肽库要求。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.

关 键 词：抗体库 合成 基因工程 肽库 

分 类 号：Q782[生物学—分子生物学]