Description: NZYTech has designed different strategies to enhance the production of difficult-to-express proteins. With that in mind, we developed a battery of E. coli expression vectors – pHTP series (included on the NZYEasy Cloning & Expression System) – which allow testing expression of any protein in fusion with several tags known to enhance solubility and/or promote correct folding. To overcome solubility problems of challenging proteins, we offer a service that includes a small-scale expression screening stage (stage 1), followed by a second stage for large-scale protein production (see table above).
The first stage includes: (1) sub-cloning of the target gene into 10 different pHTP expression vectors; (2) small-scale protein production in different expression conditions; and (3) protein expression evaluation. The second stage is devoted to produce higher amounts of the desired protein after selecting the best expression conditions.
Features:
Stage 1 – Small-scale expression screening:
– Gene amplification through PCR (or by gene synthesis at an additional price)
– Cloning into 10 pHTP E. coli expression vectors containing different tags known to enhance protein’s solubility and/or promote correct protein’s folding
– QC: verification of correct plasmids by Sanger sequencing
– Small-scale protein production trials: protein expression in different conditions – may include testing growth in LB or auto-induction media; different E. coli expression strains, and/or different induction temperatures
– Recombinant protein purification through Immobilized Metal Affinity Chromatography (IMAC)
– SDS gel analysis and protein quantification
– Protein Expression Report – protein expression evaluation
– Absolute confidentiality is guaranteed
Stage 2 – Large-scale protein production:
– Scale-up protein expression using optimized conditions
– Protein purification to reach desired protein amounts and degree of purity
– SDS-PAGE analysis and protein quantification
– Protein Production Report
– Absolute confidentiality is guaranteed
Delivery Specifications:
– Recombinant genes cloned into pHTP expression vectors (stage 1)
– Purified recombinant proteins are provided in a buffered solution with specified concentration & purity (stage 2)
– Protein Production Reports containing QC certificate, recombinant DNA sequences, recombinant protein sequences, SDS-PAGE analysis and protein quantification
– In case of not being possible to proceed with stage 2, all charges related to stage 1 will still be applicable
– Estimated delivery: upon request
Quotes & Ordering:
Please contact us to services@nzytech.com.
