Efficient Generation of Genome-wide Libraries for Protein-ligand Screens Using Gibson Assembly.

ABSTRACT: Genome-wide screens using yeast or phage displays are powerful tools for identifying protein-ligand interactions, including drug or vaccine targets, ligand receptors, or protein-protein interactions. However, assembling libraries for genome-wide screens can be challenging and often requires unbiased cloning of 10 ⁵ -10 ⁷ DNA fragments for a complete representation of a eukaryote genome. A sub-optimal genomic library can miss key genomic sequences and thus result in biased screens. Here, we describe an efficient method to generate genome-wide libraries for yeast surface display using Gibson assembly. The protocol entails genome fragmentation, ligation of adapters, library cloning using Gibson assembly, library transformation, library DNA recovery, and a streamlined Oxf

SUBMITTER: Sternlieb T 

PROVIDER: S-EPMC9724014 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature