中文摘要：

脂质运载蛋白（lipocalin）是一类广泛存在于生物界的小分子蛋白质家族,具备一些抗体所没有的优点.为了构建库容量大、多样性好的核糖体展示模拟抗体Anticalin文库,本研究以脂质运载蛋白家族之一大菜粉蝶（Pieris brassicae）中提取到的胆汁三烯结合蛋白基因（bilin-binding protein, bbp）（GenBank登录号：X76568.1）为模板,设计了包含16个随机突变位点的引物,采用重叠延伸PCR技术（gene splicing by overlap extension PCR, SOE-PCR）构建bbp基因文库.然后,利用SOE-PCR法引入核糖体展示元件（T片段与P片段）,体外构建核糖体展示模拟抗体Anticalin基因文库.结果表明,本研究成功构建了模拟抗体Anticalin基因文库,其库容达到3.76×1023,文库的可扩增性、完整性及多样性良好,为筛选多种小分子化学残留的模拟抗体提供了基础资料.

英文摘要：

Lipocalin, a family of diverse proteins that distribute widely in organisms, exhibits several advantages, compared with antibody. To construct a high-capacity and good-diversity ribosome display anticalin library for selecting small molecular anticalin with high affinity, bilin-binding protein gene（bbp） that is a member of lipocalin family from Pieris brassicae, was employed for the preparation of a random library. A DNA library, named bbp gene library, encoding amino acids of bbp possessing 16 randomly mutated residues, was generated using gene splicing by overlap extension PCR （SOE-PCR）. Then, using the same method the elements （T and P fragments） that were necessary for efficient transcription and translation in vitro were introduced to bbp gene library to construct the ribosome display anticalin library. In conclusion, we successfully constructed a ribosome display anticalin library with a volume of 3.76×1023, high-capacity and good-diversity. This library could be served as an efficient selection system for antibodies of micro-molecular haptens.