利用核糖体展示文库初步筛选与伤寒杆菌Ⅳ型纤毛结合的细胞受体

Selecting cell receptor of type Ⅳ pili in Salmonella typhi by ribosome display library

伤寒杆菌病原毒力岛上的Ⅳ型纤毛与伤寒杆菌的致病性密切相关,但对Ⅳ型纤毛在人巨噬细胞、树突状细胞上的受体及其序列或结合的关键部位还一无所知。本研究拟用核糖体展示文库筛选伤寒杆菌结合的细胞受体。体外人工合成引物和含36个随机序列的寡核苷酸,通过两轮PCR构建随机DNA文库,随后在体外进行偶联的转录和翻译,得到含随机12肽的核糖体文库。利用含随机12肽的核糖体文库与体外重组表达的Ⅳ型纤毛结构蛋白(PilS蛋白)进行了初步筛选,为筛选到与伤寒杆菌毒力岛上Ⅳ型纤毛结构蛋白结合的细胞受体奠定了基础。

TypeⅣpili in the pathogenicity island of Salmonella yyphi is relative to the pathogenicity of Salmonella ty phi ,but the cell receptor of typeⅣpili in Salmonella typhi is unknown.In this study,ribosome display library was used to select cell receptor of typeⅣpili,36random-sequence oligonucleotides and primers were designed and synthesed.A DNA random library was constructed by PCR using a random-sequence oligonucleotides template.Then a coupled tran-scription/translation reaction in vitro was proceeded by use of E.coli S30Extract System.Finally,a ribosome display li-brary including12random peptides was constructed and used to select the cell receptor of typeⅣB pili in Salmonella typhi .This study,ribosome display library including12random peptides was constructed successfully and used to screen the cell receptor of typeⅣB pili in Salmonella typhi .This was useful for the further work.