Table 1. Pichia pastoris strains generated for systematic investigation of the relationship between protein complexity, gene dosage, relative expression and secretion.
Strain Namea | Secreted protein | Gene copyb number | Relative mRNAc Expression |
pGAPαeGFP1 | eGFP | 1 | 2.1 |
pGAPαeGFP2 | eGFP | 3 | 12.7 |
pGAPαeGFP3 | eGFP | ≥4 | 19.4 |
pGAPαGFc1 | Glycosylated Fc | 1 | 1 |
pGAPαGFc2 | Glycosylated Fc | 2 | 1.4 |
pGAPαGFc3 | Glycosylated Fc | ≥6 | 11.3 |
pGAPαAgFc1 | Aglycosylated Fc | 1 | 5.1 |
pGAPαAgFc2 | Aglycosylated Fc | 2 | 5.4 |
pAOXαeGFP1 | eGFP | 2 | 16.4 |
pAOXαeGFP2 | eGFP | 5 | 19.8 |
pAOXαGFc1 | Glycosylated Fc | 2 | 10.9 |
pAOXαGFc2 | Glycosylated Fc | 5 | 8.9 |
pAOXαAgFc1 | Aglycosylated Fc | 2 | 5.1 |
pAOXαAgFc1 | Aglycosylated Fc | 4 | 22.2 |
Strain names indicate promoter used.
Determined by qPCR using an absolute quantification of transcript copy number.
Determined by RT-qPCR using an absolute quantification of transcript copy number and the expression of actin as a control.