An efficient ORF selection system for DNA fragment libraries based on split beta-lactamase complementation.

ABSTRACT: PCR-based amplification of annotated genes has allowed construction of expression clones at genome-scale using classical and recombination-based cloning technologies. However, genome-scale expression and purification of proteins for down-stream applications is often limited by challenges such as poor expression, low solubility, large size of multi-domain proteins, etc. Alternatively, DNA fragment libraries in expression vectors can serve as the source of protein fragments with each fragment encompassing a function of its whole protein counterpart. However, the random DNA fragmentation and cloning result in only 1 out of 18 clones being in the correct open-reading frame (ORF), thus, reducing the overall efficiency of the system. This necessitates the selection of correct ORF before expressi

SUBMITTER: Verma V 

PROVIDER: S-EPMC7377443 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature