Rapid screening of high expressing Escherichia coli colonies using a novel dicistronic-autoinducible system.

ABSTRACT:

Background

Identification of high-expressing colonies is one of the main concerns in the upstream process of recombinant protein development. The common method to screen high-producing colonies is SDS-PAGE, a laborious and time-consuming process, which is based on a random and qualitative way. The current study describes the design and development of a rapid screening system composed of a dicistronic expression system containing a reporter (enhanced green fluorescent protein, eGFP), protein model (staphylokinase, SAK), and a self-inducible system containing heat shock protein 27 (Hsp27).

Results

Dicistronic-autoinducible system expressed eGFP and SAK successfully in 5-ml and 1-L culture volumes. High expressing colonies were identified during 6 h via fluorescent signals. In

SUBMITTER: Shariati FS 

PROVIDER: S-EPMC8666062 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature