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Yeast Expression

      Yeast protein expression system is a ideally suitable expression system for both secretion and intracellular expression as the yeast offer relatively easy genetic manipulation and rapid growth to high cell densities on inexpensive media. Common yeast protein expression systems are Pichia pastoris, Saccharomyces cerevisiae, Kluyveromyces lactis, Hansenula polymorpha, and Yarrowia lipolytica. TGpeptide use Pichia pastoris as our strain.

Service procedures

Specification

Timeline

1. Analysis of gene and target protein

Codon optimization, protein hydrophilic, hydrophobic, PI, structural domain, etc

1-2 days

2.Subcloning

Gene synthesis

Construction of expression vector

Plasmids Prep and analysis

3.Screening of expression strains

Amplification of recombinant plasmids and linear electrotransformation of Pichia pastor with the recombinant plasmids

Obtain high-copy strain by antibiotic screening

Small amount expression with ten selected positive transformation strains and protein expression evaluation and optimization

SDS-PAGE to identify the expression

3-4 weeks

4.Protein expression and purification in large scale

Large amount expression with selected strains of high-expression

Protein purification

SDS-PAGE test

2-3 weeks

5.Screening of expression strains

High density cultivation of protein expression in fermentation tanks and purification

Carry out the experiment with the optimized expression condition

Protein purification SDS-PAGE test

3-4 weeks

Deliverables:

Expression vector containing the target gene

Target sequencing report.

E.coli containing plasmids of the expression vector

Plasmid instructions

SDS-PAGE analysis results

Expression strain (extra charge, $1500 / strain)

Purified recombinant protein (purity >90%)

Related analysis reports

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