E. Coli Expression System

 

Escherichia coli is one of the most widely used hosts for the heterologous protein expression. Its genetics are far better characterized than those of any other microorganism.

 

Service description

 

All services include multiple steps from gene to purified protein as described in the table below.

Description Resulting Material Average turnaround (*)
Gene Synthesis Stage
-Codon optimization
-Gene synthesis
-Cloning in standard pUC plasmid
-Complete sequencing
Cloned, synthesized and verified gene 1-2 weeks
Subcloning in Expression Vector Stage
-Plasmid prep
-Subcloning in expression vector
-Clone selection
-Complete sequencing
Expression vector with cloned target gene 1-2 weeks
Pilot Expression Stage
-Expression vector prep
-Transformation using 3 different strains
-Protein induction and optimzation experiments
-Determine yields and protein behavior
10-50 µg of semi-purified protein 1-2 weeks
Expression and Purification Stage
-Expression of 1 to 5 L bacterial culture
-Protein extraction
-Protein purification
-SDS-PAGE, WESTERN blot
1-10 mg of purified protein
Purity >80% or >90%
2-3 weeks

(*) Average production time for average difficult project. Actual turn around time varies with individual project features.

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