TABLE 5.
Strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source |
|---|---|---|
| Strains | ||
| Thanatephorus cucumeris NBRC 6298 | Wild type | NBRC |
| Pichia pastoris X33 | Wild type | Invitrogen |
| Escherichia coli TOP 10 | F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu) 7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| Pichia pastoris X33-CK | Pichia pastoris X33 harboring pICZA empty vector | This work |
| Pichia pastoris X33-POR | Pichia pastoris X33 expressing por alone | This work |
| Pichia pastoris X33-STH10 | Pichia pastoris X33 expressing sth10 alone | This work |
| Pichia pastoris X33-STH10-POR | Pichia pastoris X33 coexpressing sth10 and por | This work |
| Plasmids | ||
| pPICZ A | Yeast expression vector (Pichia pastoris), Zeor | Invitrogen |
| pUC57 | Cloning vector, Ampr | Invitrogen |
| pPICZ-STH10 | T. cucumeris sth10 cDNA cloned in pPICZ A vector at KpnI-NotI site; Zeor | This work |
| pUC57-POR | T. cucumeris P450 reductase gene por cloned in pUC57 vector at KpnI-NotI site; Ampr | This work |
| pPICZ-MPOR | pPICZ A-por vector containing por gene mutated (silent mutation) at EcoRI; Zeor | This work |
| dePmeI-pPICZA | pPICZ A vector mutated at PmeI site; Zeor | This work |
| binary-pPICZ-MPOR | BglII-BamHI digested dePmeI-pPICZ A fragment cloned in pPICZ-MPOR at BamHI site; Zeor | This work |
| pPICZ-STH10-POR | sth10 gene cloned in binary-pPICZ-MPOR at KpnI-NotI site; Zeor | This work |
a
Ampr, ampicillin resistance; Strr, streptomycin resistance; Zeor, zeocin resistance.