Table 1 Plasmids and strains used in this work
Plasmids/strains | Genotype | Reference |
|---|---|---|
Plasmids | ||
pGAPZB | Pichia integrative plasmid; Zeocin® selection; Expression controlled by constitutve GAP promoter | Invitrogen® |
pPICPGKGFP | Green Fluorescente enconding gene cloned under the controlo f PGK promoter | Personal communication |
pGAP-LDH | LDH+, Bos taurus gene encoding for LDH enzyme | This work |
pPGK-JEN | JEN1+, S. cerevisiae gene-encoding for the lactate transporter Jen1p | This work |
pPGK-PAS | PAS+, P. pastoris gene-encoding for a putative lactate transporter PAS | This work |
E. coli strains | ||
DH5α™ | F– Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK–, mK +) phoA supE44 λ– thi-1 gyrA96 relA1 | Life technology |
DH10B™ | F– mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15ΔlacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– | Life technology |
P. pastoris strains | ||
X-33 | Wild type | Life technology |
XL | X-33 + pGAP-LDH | |
GS115 | his4 − | Life technology |
GJ | his4− + pPGK-JEN | This work |
GLJ | his4− + pGAP-LDH + pPGK-JEN | This work |
GS | his4− + pPGK-PAS | This work |
GLS | his4− + pGAP-LDH + pPGK-PAS | This work |