Simplified cloning and isolation of peptides from "sandwiched" SUMO-peptide-intein fusion proteins.

ABSTRACT:

Background

Some peptides are targets for degradation when heterologously expressed as fusion proteins in E. coli, which can limit yields after isolation and purification. We recently reported that peptide degradation may be prevented by production of a "sandwiched" SUMO-peptide-intein (SPI) fusion protein, which protects the target peptide sequence from truncation and improves yield. This initial system required cloning with two commercially available vectors. It used an N-terminal polyhistidine tagged small ubiquitin-like modifier (SUMO) protein and a C-terminal engineered Mycobacterium xenopii DNA Gyrase A intein with an inserted chitin binding domain (CBD) to create "sandwiched" fusion proteins of the form: His₆-SUMO-peptide-intein-CBD. However, the major drawback of

SUBMITTER: Lamer T 

PROVIDER: S-EPMC10074672 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature