EXPERIMENTS
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Escherichia coli vectors
pET28
The pET28 vector contains two His Tags in his MCS and is specified for protein expression at high yield. It give the strain a resistance in Kanamycin and is replication origin is the pBR322 which is a mid - low copy number plasmid (~10). The vector used was a kind gift from IBCG library.
pETDuet-1
The pETDuet-1 is, as the pET28, specialized in high yield protein production. It main difference is that it contains two MCS and can therefore express two proteins at the same time under the same expression conditions. It bears ampicilin resistance and is a mid - low copy number plasmid. The plasmid used was a gift from IBCG library.
Pichia pastoris vectors
pGAPZ alpha
The pPICZalpha i an integrative plasmid for Pichia pastoris protein expression and suitable for E. coli replication. Its MCS is suitable for protein expession using the methanol inductible AOX1 Pomoter (PAOX1) and offering the alpha factor secretion signal for protein purification from culture supernatant. It contains a His tag and a Myc Tag for affinity purification stategies.
pET28
The pGAPZalpha is roughly the same as the pPICZalpha with the difference of the promoter that is GAP Promoter, a glucose indcutible promoter.
Escherichia coli strains
Plasmid amplification strains- Stellar
- Top10
- Stellar
- Top10
Pichia pastoris strains
For protein production we used GS200 Strain for Cerberus and X33 for Scygonadin
- DBCO-Biotin
- DBCO-Fluorescein
- DBCO-Magnetic beads
No dogs were harmed over the course of this iGEM project.
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