Increasing recombinant protein production in E. coli via FACS-based selection of N-terminal coding DNA libraries.

ABSTRACT: Here, we present a previously undescribed approach to modify N-terminal sequences of recombinant proteins to increase their production yield in Escherichia coli. Prior research has demonstrated that the nucleotides immediately following the start codon can significantly influence protein expression. However, the impact of these sequences is construct-specific and is not universally applicable to all proteins. Most of the previous research has been limited to selecting from a few rationally designed sequences. In contrast, we used a directed evolution-based methodology, screening large numbers of diversified sequences derived from DNA libraries coding for the N-termini of investigated proteins. To facilitate the identification of cells with increased expression of the target construct, we c

SUBMITTER: Herynek S 

PROVIDER: S-EPMC11880969 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature