Accelerated High Fidelity Prion Amplification Within and Across Prion Species Barriers 英文参考文献.docVIP

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Accelerated High Fidelity Prion Amplification Within and Across Prion Species Barriers 英文参考文献.doc

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Accelerated High Fidelity Prion Amplification Within and Across Prion Species Barriers 英文参考文献

AcceleratedHighFidelityPrionAmplificationWithinand AcrossPrionSpeciesBarriers KristiM.Green1¤a,Joaqu?′nCastilla2¤b,TanyaS.Seward3,DanaL.Napier3,JeanE.Jewell4,ClaudioSoto2, GlennC.Telling1,3,5 * 1DepartmentofMicrobiology,ImmunologyandMolecularGenetics,UniversityofKentucky,Lexington,Kentucky,UnitedStatesofAmerica,2DepartmentofNeurology, UniversityofTexasMedicalBranch,Galveston,Texas,UnitedStatesofAmerica,3SandersBrownCenteronAging,UniversityofKentucky,Lexington,Kentucky,United StatesofAmerica,4DepartmentofVeterinarySciences,UniversityofWyoming,Laramie,Wyoming,UnitedStatesofAmerica,5DepartmentofNeurology,Universityof Kentucky,Lexington,Kentucky,UnitedStatesofAmerica Abstract Experimentalobstacleshaveimpededourabilitytostudypriontransmissionwithinand,moreparticularly,betweenspecies. Here,weusedcervidprionproteinexpressedinbrainextractsoftransgenicmice,referredtoasTg(CerPrP),asasubstrate for in vitro generation of chronic wasting disease (CWD) prions by protein misfolding cyclic amplification (PMCA). Characterization of this infectivity in Tg(CerPrP) mice demonstrated that serial PMCA resulted in the high fidelity amplificationofCWDprionswithapparentlyunalteredproperties.UsingsimilarmethodstoamplifymouseRMLprionsand characterize the resulting novel cervid prions, we show that serial PMCA abrogated a transmission barrier that required several hundred days of adaptation and subsequent stabilization in Tg(CerPrP) mice. While both approaches produced cervidprionswithcharacteristicsdistinctfromCWD,thesubtlydifferentpropertiesoftheresultingindividualprionisolates indicatedthatadaptationofmouseRMLprionsgeneratedmultiplestrainsfollowinginter-speciestransmission.Ourstudies demonstrate that combined transgenic mouse and PMCA approaches not only expedite intra- and inter-species prion transmission,butalsoprovideafacilemeansofgeneratingandcharacterizingnovelprionstrains. Citation: Green KM, Castilla J, Seward TS, Napier DL, Jewell JE, et al. (2008) Accelerated High Fidelity Prion