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Custom Protein Services:  Yeast Expression Systems

Insight Biotechnology provides a broad range of yeast protein expression services which can be tailored to your specific research needs.  Choose any starting phase or combination.   Insight Biotechnology takes a flexible approach towards custom protein services and information on this website is only a guide.  Feel free to contact our team of protein expression experts with any further questions.   Email, or telephone Insight Biotechnology on Tel:  +44(0)208 3850303.  


Please Click here for further information about other custom services offered by Insight Biotechnology.

Phase 1: Subcloning into Expression Vector

Description

Amplification/isolation of the gene of interest out of a customer-supplied vector and subcloning it into a suitable expression vector.

 

Verification of the subcloned gene by restriction digest and sequencing

 

Preparation of the recombinant vector DNA.

 

 

Required Materials

5µg of plasmid containing gene of interest plus complete vector sequence in electronic format.

 

 

Deliverables

1) 20µg of expression-ready recombinant plasmid containing the gene of interest

 

2) Final sequencing data

 

3) Detailed research report

 

 

Duration

2 weeks; If insert > 2kb, one week additional per 1kb

 

 

Price

Enquire

 

 


Phase II: Generation and Identification of Expression Positive Clones

Description

Transformation of recombinant constructs into a suitable yeast strain.

 

Small scale induction to over-express target protein.

 

Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies).

 

 

Required Materials

5µg of expression-ready recombinant plasmid containing the gene of interest plus complete vector sequence in electronic format.

 

 

Deliverables

1) Expression positive clones ( E. coli glycerol stock )

 

2) SDS-PAGE and Western Blot data

 

3) Detailed research report

 

 

Duration

2 weeks

 

 

Price

Enquire

 

 

Optional

For His or GST tagged protein, you can choose to use our antibodies in stock, with an additional charge of £20.


Phase III: Large-scale Culture

Description

One litre of yeast culture will be grown in induction medium suitable for the expression vector used. Recombinant protein expression is harvested for protein purification.

 

Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies).

 

 

Required Materials

Expression positive yeast clone together with a verified induction procedure

 

 

Deliverables

1) Cell pellets from 1 litre fermentation

 

2) SDS-PAGE or Western Blot data

 

3) Detailed research report

 

 

Duration

1 Week

 

 

Price

Enquire

 

 

Optional

For His or GST tagged protein, you can choose to use our antibodies in stock, with an additional charge of £20.


Phase IV: Protein Extraction & Purification

Description

Cell pellet harvested from 1 Litre of yeast culture after induction will be lysed with our optimised reagents and protocol.

 

Protein purification using appropriate affinity resin for 6xHis or GST-tagged recombinant proteins.

 

Purification can be performed either under native or denatured conditions (as desired by the customer).

 

SDS Page and Western Blot will be performed for quality testing.

 

 

Required Materials

Cell pellet harvested from 1 litre of bacterial after induction

 

 

Deliverables

1) Purified protein in standard or customer-specified solution

 

2) SDS-PAGE or Western Blot data

 

3) Detailed research report

 

 

Duration

2-3 weeks

 

 

Price

Enquire

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