Hale M B, Nolan G P, Wolkowicz R
Department of Molecular Pharmacology, School of Medicine, Stanford University, Stanford, CA 94305, USA.
Nucleic Acids Res. 2004 Jan 29;32(2):e22. doi: 10.1093/nar/gnh021.
We present an approach that generates an oligomer-based library with minimal need for restriction site modification of sequences in the target vector. The technique has the advantage that it can be applied for generating peptide aptamer libraries at sites within proteins without the need for introducing flanking enzyme sites. As an example we present a phagemid retroviral shuttle vector that can be used to achieve stable expression of the library in mammalian cells for the purpose of screening for peptides with desired biological activity.
我们提出了一种方法,该方法生成基于寡聚物的文库,对靶载体中序列的限制性酶切位点修饰需求最小。该技术的优点是可应用于在蛋白质内部位点生成肽适配体文库,而无需引入侧翼酶切位点。作为一个例子,我们展示了一种噬菌粒逆转录病毒穿梭载体,其可用于在哺乳动物细胞中实现文库的稳定表达,以筛选具有所需生物活性的肽。