利用烟草基因组DNA构建近随机多肽文库

介绍一种新的方法构建近随机多肽文库。选取从大基因组物种的组织或细胞中提取的基因组DNA ,利用切割频率高的限制性内切酶切割 ,产生的短片段可以近似地认为是随机序列的片段 ,将它们与匹配的载体连接后转化进宿主细胞进行表达 ,从而获得近随机多肽文库。这样的文库可以用于蛋白质相互作用的研究。同一种基因组DNA可以利用不同的酶切 ,再分别连接到表达载体的不同读码框架 ,从而产生不同编码序列的多种近随机多肽文库。介绍了充分利用烟草基因组DNA构建两种不同酶切 ,三种读码框架 ,共六种不同编码序列的近随机多肽文库的方法。

Utilizing Tabacco Genomic DNA to Construct Nearly Random Peptide Libraries

We developed a novel method for constructing nearly rand om peptide library. Genomic DNAs extracted from tissue or cells of large genome species were digested with frequent cutter to produce short DNA fragments. These short fragments can be considered nearly random. Nearly random peptide librarie s can be constructed by cloning the short fragments into appropriate expression vectors and transformation into host cells. Genomic DNA from one species can be digested with different restriction enzymes and ligated to different reading fra mes to produce several different libraries. In this study, we digested tobacco g enomic DNA with two enzymes and cloned into three different reading frames to ma ke totally six nearly random peptide libraries.