Prokaryotic expression system (Escherichia coli expression system) is the earliest developed and most widely used classical expression system in gene expression technology. In recent decades, Escherichia coli expression system has been continuously developed and improved, and has been widely used for various recombinant protein expression by scientific research and industrial users. Compared with other expression systems, it has the characteristics of high expression level of target gene, short culture cycle, strong anti-pollution ability and relatively low cost.

1. scheme design - analysis of target gene sequence, according to customer requirements and existing literature reports to make experimental scheme 

services Our company provides free services for customers: gene codon optimization, protein signal peptide analysis, protein hydrophobicity analysis, protein transmembrane analysis, etc.

2. Construction of recombinant plasmids within 1-2 weeks. Appropriate vectors and appropriate labels (eg: HIS, TRX, GST, SUMO)are selected to construct recombinant plasmids, supporting site-directed mutagenesis, vector modification, seamless cloning and other services. The company can provide free of charge: PET series intracellular or secretory expression vectors, PGEX series intracellular expression vectors; Thermo-controlled PBV220 intracellular or secretory expression vectors.

3. Induced expression of recombinant protein within 1 week, appropriate methods are selected to induce the expression of recombinant protein, and SDS-PAGE is used to detect the expression of protein, and the expression mode of protein is detected as soluble expression or inclusion body reach.

4. Separation and purification of recombinant proteins within 1 week. The most commonly used chromatographic methods for separation and purification of recombinant proteins are: affinity chromatography (Ni, GST), anion and cation exchange chromatography (Q, SP), molecular exclusion chromatography, etc.

5. QC analysis of the recombinant protein within.1 week SDS-PAGE electrophoresis is used to detect the purity of the recombinant protein, the concentration of the recombinant protein is quantified by BCA, and the immunogenicity of the recombinant protein is tested by Western-blot.

Recombinant protein reprocessing for 1 week (optional, extra charge is required) Recombinant protein filtering and degerming, endotoxin removal, label cutting, etc.