TABLE 5

Strains and plasmids used in this study

Strain or plasmidGenotype or phenotypeSource or reference
Strains
 Thanatephorus cucumeris NBRC 6298Wild typeNBRC
 Pichia pastoris X33Wild typeInvitrogen
 E. coli TOP 10F mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu) 7697 galU galK rpsL (StrR) endA1 nupGInvitrogen
    X33-CKPichia pastoris X33 harboring pICZA empty vector21
    X33-PORPichia pastoris X33 expressing por alone21
    X33-CYP5150AP3Pichia pastoris X33 expressing cyp5150ap3 aloneThis work
    X33-CYP5150AN1Pichia pastoris X33 expressing cyp5150an1 aloneThis work
    X33-CYP5150AP3-PORPichia pastoris X33 coexpressing cyp5150ap3 and porThis work
    X33-CYP5150AN1-PORPichia pastoris X33 coexpressing cyp5150an1 and porThis work
Plasmids
    pPICZAYeast expression vector (Pichia pastoris); ZeorInvitrogen
    pUC57Cloning vector; AmprInvitrogen
    pPICZ-CYP5150AP3T. cucumeris cyp5150ap3 cDNA cloned in pPICZA vector at KpnI-NotI site; ZeorThis work
    pPICZ-CYP5150AN1T. cucumeris cyp5150an1 cDNA cloned in pPICZA vector at KpnI-NotI site; ZeorThis work
    pUC57-PORT. cucumeris P450 reductase gene por cloned in pUC57 vector at KpnI-NotI site; Ampr21
    pPICZ-MPORpPICZA-por vector containing por gene mutated (silent mutation) at EcoRI; Zeor21
    dePmeI-pPICZApPICZA vector mutated at PmeI site; Zeor21
    binary-pPICZ-MPORBglII-BamHI-digested dePmeI-pPICZA fragment cloned in pPICZ-MPOR at BamHI site; Zeor21
    pPICZ-CYP5150AP3-PORcyp5150ap3 gene cloned in binary-pPICZ-MPOR at KpnI-NotI site; ZeorThis work
    pPICZ-CYP5150AN1-PORcyp5150an1 gene cloned in binary-pPICZ-MPOR at KpnI-NotI site; ZeorThis work