First Discovery of Two Polyketide Synthase Genes for Mitorubrinic Acid and Mitorubrinol Yellow Pigment Biosynthesis and Implications in Virulence of Penicillium marneffei 英文参考文献.docVIP

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First Discovery of Two Polyketide Synthase Genes for Mitorubrinic Acid and Mitorubrinol Yellow Pigment Biosynthesis and Implications in Virulence of Penicillium marneffei 英文参考文献.doc

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First Discovery of Two Polyketide Synthase Genes for Mitorubrinic Acid and Mitorubrinol Yellow Pigment Biosynthesis and Implications in Virulence of Penicillium marneffei 英文参考文献

FirstDiscoveryofTwoPolyketideSynthaseGenesfor MitorubrinicAcidandMitorubrinolYellowPigment BiosynthesisandImplicationsinVirulenceofPenicillium marneffei PatrickC.Y.Woo1,2,3,4*.,Ching-WanLam5.,EmilyW.T.Tam1.,ChrisK.F.Leung5, SamsonS.Y.Wong1,2,3,4,SusannaK.P.Lau1,2,3,4*,Kwok-YungYuen1,2,3,4 1StateKeyLaboratoryofEmergingInfectiousDiseases,TheUniversityofHongKong,HongKong,2ResearchCentreofInfectionandImmunology,TheUniversityofHong Kong,HongKong,3CarolYuCentreforInfection,TheUniversityofHongKong,HongKong,4DepartmentofMicrobiology,TheUniversityofHongKong,HongKong, 5DepartmentofPathology,TheUniversityofHongKong,HongKong Abstract Background: The genome of P. marneffei, the most important thermal dimorphic fungus causing respiratory, skin and systemic mycosis in China and Southeast Asia, possesses 23 polyketide synthase (PKS) genes and 2 polyketide synthase nonribosomalpeptidesynthasehybrid(PKS-NRPS)genes,whichisofhighdiversitycomparedtootherthermaldimorphic pathogenicfungi.WehypothesizedthattheyellowpigmentinthemoldformofP.marneffeicouldalsobesynthesizedby oneormorePKSgenes. Methodology/PrincipalFindings:All23PKSand2PKS-NRPSgenesofP.marneffeiweresystematicallyknockeddown.A loss of the yellow pigment was observed in the mold form of the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants.Sequence analysis showed that PKS11 andPKS12 are fungal non-reducing PKSs. Ultra high performance liquid chromatography-photodiode array detector/electrospray ionization-quadruple time of flight-mass spectrometry (MS) andMS/MS analysis of theculture filtratesof wild type P.marneffei and thepks11 knockdown, pks12 knockdownandpks11pks12doubleknockdownmutantsshowedthattheyellowpigmentiscomposedofmitorubrinicacid andmitorubrinol. Thesurvivalofmice challenged withthe pks11 knockdown, pks12knockdown andpks11pks12 double knockdown mutantswas significantly better thanthose challenged withwild type P. marneffei (P,0.05). There was also statistically significant decrease in